Inositol derivatives for increasing chloride secretion and inhibiting inflammation

ABSTRACT

Inositol derivatives, compositions comprising inositol derivatives, and methods for using compositions comprising inositol derivatives as agents for activating the secretion of chloride ions and/or treatment of inflammation are described.

FIELD OF THE INVENTION

[0001] The present invention relates to inositol derivatives thatactivate the secretion of chloride ions, promoting hydration of mucosalsurfaces which is deficient in mucosal epithelia of patients sufferingfrom cystic fibrosis, and/or that inhibit inflammation. The presentinvention also relates to compositions comprising the inositolderivatives and methods for their use, alone or in combination withother therapeutic agents, for treating pathological conditions relatedto cystic fibrosis and/or inflammation in humans.

BACKGROUND OF THE INVENTION

[0002] Cystic fibrosis(CF) is the most common genetic disorder and thelargest genetic killer of children. One in twenty Caucasians carries adefective CF gene, which, when coupled with a spouse who is also acarrier can result in offspring afflicted with CF. An autosomal,recessive disorder, one in 3,000 children born in the United States andEurope inherit CF. Children live for varying periods of time, but theaverage has been extended from a couple of years early in this centuryto a current life expectancy of 30 years. Over 70,000 patients have beenidentified with Cystic Fibrosis worldwide. This translates into over30,000 individuals with the disease in the United States with another30,000 who have been identified with the disorder in Europe. As currenttreatment strategies prolong the average lifespan, the number of CFpatients is expected to rise. Patients with CF typically incur medicalcosts ranging from $15,000 to $55,000 annually. In addition, carriers ofCF (about 5% of Caucasians) have a higher likelihood of suffering fromsinusitis, a disease of the sinus passages characterized by recurrentinfections.

[0003] CF is due to a genetic defect in a chloride-channel in mucusmembranes that regulates the passage of chloride, sodium and water intomucous linings. This leads to unusually thick mucus, particularly in thelungs, which encourages the growth of bacteria and consequently repeatedinfections and inflammation.

[0004] In a majority of CF victims, the cystic fibrosis transmembraneregulator (CFTR), an epithelial chloride channel, is defective. In 70%of the cases this is due to a single point mutation at ΔF508. Thus, CFmucosal epithelial cells are incapable of transferring sufficientamounts of chloride ions to the outer membrane surface to draw enoughwater to adequately hydrate the mucous membranes. The inherited diseaseprocess of CF affects tissues of mucous membranes such as those liningthroat, nasal, sinus, bronchial passages, lungs, gall bladder, sweatglands, pancreas and intestines.

[0005] The disease causes abnormally viscous mucous secretions that leadto chronic pulmonary disease, pancreatic insufficiency and intestinalobstructions, together with a host of lesser but potentially lethalproblems, such as an excessive loss of electrolytes in hot environments.In the past, afflicted children often died as infants. Althoughsurviving into their twenties and thirties with current treatments, CFpatients are plagued with recurrent infections and require daily arduousroutines to clear air passageways.

[0006] It is believed that agents that promote chloride secretion suchas purinergic agonists and other agents that elevate intracellularcalcium could be useful in increasing hydration of the airways, thusreducing mucous viscosity. However, it is known that nucleotides alreadypresent in the fluid lining the lungs should be sufficient to stimulatea rise in levels of intracellular calcium. Thus, it may also benecessary to inactivate pathways that limit chloride secretion, such asthose mediated by inositol polyphosphate and phosphoinositide pathwaysdescribed below.

[0007] It has recently been shown that certain inositol phosphatederivatives including D-myo-inositol 3,4,5,6-tetrakisphosphate(“Ins(3,4,5,6)P₄”) (M. Vajanaphanich et al., Nature 371:711 (1994); J.M. Uribe et al., J. Biol. Chem. 271(43):26588 (1996); M. T. Rudolf etal., J. Med. Chem. 41 (19):3635-3644 (1998)), andsn-di-O-palmitoylD,L-6-O-butyryl-phosphatidylinositol3,4,5-trisphosphate heptakis(acetoxymethyl)ester (C. Schultz et al.,Membrane-permeant, Bioactivatable Derivatives of Inositol Polyphosphatesand Phosphoinositides, in Phosphoinositides. Chemistry, Biochemistry andBiomedical Applications, K. S. Bruzik, Ed. Am. Chem. Soc., Symp. Ser.,718, 232-243 (1999)) inhibit calcium-mediated chloride secretion.D-myo-Inositol 1,4,5,6-tetrakisphosphate has been shown to inhibit aphosphatidylinositol PI-3 kinase (“PI-3 kinase”) signaling pathway incolonic epithelia (Eckmarn et al., Proc. Natl. Acad. Sci. USA 94:14456(1997)). It has also recently been shown that2,6-di-O-butyryl-myo-inositol 1,2,4,5-octakis(acetoxymethyl)esterincreased the level of Ca²⁺ in PC12 cells (C. Schultz et al. (1998),supra). U.S. Pat. No. 5,693,521 to Tsien et al. discloses the use ofD-2,3-di-O-butyryl-myo-inositol 1,4,5,6-tetrakisphosphateoctakis(acetoxy-methyl)ester as a second messenger having enhanced cellpermeability. International Publication No. WO 98/11901 byTraynor-Kaplan et al. describes various inositol phosphates withenhanced cell permeability that function as either agonists orantagonists of inositol polyphosphates. In addition, the synthesis ofother inositol phosphates has been reported (S. Roemer et al., J. Chem.Soc.; Perkin Trans. 1, 1683 (1996); International Publication No. WO96/40695 to Tsien et al.; International Publication No. WO 98/11901 toA. Traynor-Kaplan et al.; Rudolf, M. T. et al., Bioorg. & Med. Chem.Lett., 8:1857 (1998); Jiang, T. et al., J. Biol. Chem. 273:11017 (1998);Li, W., et al., Tetrahedron 53:12017 (1997)).

[0008] Inflammation also plays a role in cystic fibrosis and otherdisorders. Inflammation is the body's response to injury and has beendefined as a localized protective response to destroy, dilute orsequester the injurious agent and/or injured tissue. Classic macroscopicsigns include pain, heat, redness, swelling and loss of function. On amicroscopic level there is a dilatation of blood vessels withaccompanying increased permeability and blood flow, exudation of fluidsand infiltration of leucocytes. These responses are triggered by therelease from the vascular system of inflammatory response mediators(e.g., histamine, prostaglandins, and leukotrienes) to destroy anybacteria or other toxins in the injured area and remove necrotic tissue.If this inflammatory response is somehow inappropriately triggered,toxic substances (i.e., reactive oxygen species-free radicals) that areassociated with the inflammatory response mediators can destroy healthytissues. A number of diseases such as asthma, osteoarthritis, rheumatoidarthritis, inflammatory bowel disease and adult respiratory distresssyndrome (ARDS) result from such a faulty inflammatory response. Inaddition, consequences of acute inflammation result in irreparabletissue damage following heart attacks and strokes. In these cases muchof the tissue destruction is caused by caustic oxygen radicals and theirbyproducts. It is now recognized that excessive release of oxygenradicals can lead to an overproduction of “wound healing” reactions suchas release of growth factors that induce overproliferation of connectivetissue and extracellular matrix components that can result in fibrosis,sclerosis, adhesions and scarring. Current therapies for theseconditions such as steroids are ameliorative and do not resolve the coreproblems and are generally immunosuppressive. In addition, the availablemedications lead to side effects that restrict their long-term use.Therefore, a need persists for safe and effective anti-inflammatoryagents.

[0009] To the best of Applicants' knowledge, the novel inositolpolyphosphate derivatives of the present invention and the use ofinositol polyphosphate derivatives for treating cystic fibrosis have notbeen previously known.

SUMMARY OF THE INVENTION

[0010] It is known that certain inositol polyphosphates elevateintracellular calcium levels, thus leading to increased chloridesecretion. In one aspect, the present invention provides novelcompounds, useful for regulating chloride secretion and/or inhibitinginflammation, having the general formula (I):

[0011] wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, and —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0012] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0013] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups;

[0014] and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.

[0015] The present invention further provides compositions, useful forregulating chloride secretion and/or inhibiting inflammation, comprisinga therapeutically effective amount of compounds of formula (I) or aracemate thereof, and pharmaceutically acceptable salts thereof,together with a pharmaceutically acceptable carrier or vehicle.

[0016] The invention further provides methods for regulating chloridesecretion and/or inhibiting inflammation in a cell or tissue with aneffective amount of a compound of formula (I), or regulating chloridesecretion and/or inhibiting inflammation in a human or animal patientcomprising administering to a patient in need of such increased chloridesecretion a therapeutically effective amount of a compound of formula(I), or a racemate thereof, or a pharmaceutically acceptable saltthereof, wherein:

[0017] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0018] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0019] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆allyl, and OC(O)C₁-C₆ all groups, including racemates and diastereomersthereof.

[0020] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (I) selected fromthe group consisting of D-2,3,5-tri-O-butyryl-myo-inositol1,4,6-trisphosphate hexakis(acetoxymethyl)ester,D-2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(propionoxymethyl) ester, and a racemate or pharmaceuticallyacceptable salt thereof.

[0021] In one embodiment of the invention, the compound of formula (I)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0022] The present invention still further provides novel compounds,useful for regulating chloride secretion and/or inhibiting inflammation,having the general formula (II):

[0023] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, and —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0024] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0025] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof, with the proviso that the compound of formula(II) is not D-3,6-Di-O-butyryl-myo-inositol 1,2,4,5-tetrakisphosphateoctakis(acetoxymethyl) ester; D-1,4-Di-O-butyryl-myo-inositol2,3,5,6-tetrakisphosphate octakis(acetoxymethyl)ester or a racematethereof.

[0026] The present invention further provides compositions, useful forregulating chloride secretion and/or inhibiting inflammation, comprisinga therapeutically effective amount of compounds of formula (II) or aracemate thereof and pharmaceutically acceptable salts thereof, with theproviso that the compound of formula (II) is notD-3,6-Di-O-butyryl-myo-inositol 1,2,4,5-tetrakisphosphateoctakis(acetoxymethyl)ester, D-1,4-Di-O-butyryl-myo-inositol2,3,5,6-tetrakisphosphate octakis(acetoxymethyl)ester, or a racematethereof. Such compositions can also comprise a pharmaceuticallyacceptable carrier or vehicle.

[0027] The invention further provides methods for regulating chloridesecretion and/or inhibiting inflammation in a cell or tissue with aneffective amount of a compound of formula (II), or regulating chloridesecretion and/or inhibiting inflammation in a human or animal patientcomprising administering to a patient in need of such increased chloridesecretion a therapeutically effective amount of a compound of formula(II), or a racemate thereof, or a pharmaceutically acceptable saltthereof, wherein:

[0028] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0029] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain allyl; and

[0030] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, —NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof. In one embodiment of the invention, the compoundof formula (II) is administered in a composition comprising apharmaceutically acceptable carrier or vehicle. Such carrier or vehiclecan include a liposomal formulation or a formulation in an aerosol.

[0031] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (II) selectedfrom the group consisting of D-3,6-di-O-butyryl-myo-inositol1,2,4,5-tetrakisphosphate octakis(propionoxymethyl)ester;D-1,4-di-O-butyryl-myo-inositol 2,3,5,6-tetrakisphosphateoctakis(propionoxymethyl)ester and a racemate or a pharmaceuticallyacceptable salt thereof.

[0032] In one embodiment of the invention, the compound of formula (II)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0033] In other embodiments, it has now been discovered that certaininositol polyphosphate derivatives act as antagonists of Ins(3,4,5,6)P₄and Ins(1,4,6)P₃, and that the Ins(3,4,5,6)P₄ antagonists can be used toactivate chloride secretion, when desired, such as in the treatment ofcystic fibrosis. The present invention still further provides novelcompounds, useful for regulating chloride secretion and/or inhibitinginflammation, having the general formula (III):

[0034] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight- or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂; or each R₁ may be taken together to form a cyclic structure;

[0035] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0036] each R₃ is independently selected from a group consisting of—C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight or branchedchain alkenyl or alkynyl, —C₁-C₁₁—X—Y, where each X is independentlyselected from a group consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—,—CO(S)—, —OC(O)—, —SC(O)—, OC(S)—, or —NH—, and where Y is independentlyselected from a group consisting of —C₁-C₁₁ straight or branched chainallyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof

[0037] each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0038] The present invention further provides compositions, useful forregulating chloride secretion and/or inhibiting inflammation, comprisinga therapeutically effective amount of compounds of formula (II) or aracemate thereof, and pharmaceutically acceptable salts thereof. Suchcompositions can also comprise a pharmaceutically acceptable carrier orvehicle.

[0039] The invention further provides methods for regulating chloridesecretion and/or inhibiting inflammation in a cell or tissue with aneffective amount of a compound of formula (III), or regulating chloridesecretion and/or inhibiting inflammation in a human or animal patientcomprising administering to a patient in need of such increased chloridesecretion a therapeutically effective amount of a compound of formula(III), or a racemate thereof, or a pharmaceutically acceptable saltthereof, wherein

[0040] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;or each R₁ may be taken together to form a cyclic structure;

[0041] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0042] each R₃ is independently selected from a group consisting of—C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight or branchedchain alkenyl or alkynyl, —C₁-C₁₁—X—Y, where each X is independentlyselected from a group consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—,—CO(S)—, —OC(O)—, —SC(O)—, OC(S)—, or —NH—, and where Y is independentlyselected from a group consisting of —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof.

[0043] each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof. In one embodiment of the invention, the compoundof formula (III) is administered in a composition comprising apharmaceutically acceptable carrier or vehicle. Such carrier or vehiclecan include a liposomal formulation or a formulation in an aerosol.

[0044] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (III) selectedfrom the group consisting of rac-(3aS, 4R, 5S, 6R, 7S,7aS)4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol; rac-(1S, 2R, 3S, 4R, 5S,6S)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane;rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[Dis(acetoxy-methoxy)phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane;or a racemate or a pharmaceutically acceptable salt thereof.

[0045] The present invention still further provides novel compounds,useful for regulating chloride secretion and/or inhibiting inflammation,having the general formula (IV):

[0046] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂;

[0047] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0048] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof; and

[0049] each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)— and where

[0050] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂, and where

[0051] n is an integer from 2 to 13, preferably 2 to 5.

[0052] The present invention further provides compositions, useful forregulating chloride secretion and/or inhibiting inflammation, comprisinga therapeutically effective amount of compounds of formula (IV) or aracemate thereof, and pharmaceutically acceptable salts thereof. Suchcompositions can also comprise a pharmaceutically acceptable carrier orvehicle.

[0053] The invention further provides, methods for regulating chloridesecretion and/or inhibiting inflammation in a cell or tissue with aneffective amount of a compound of formula (IV), or regulating chloridesecretion and/or inhibiting inflammation in a human or animal patient inneed of such increased chloride secretion a therapeutically effectiveamount of a compound of formula (IV), and racemates, stereoisomers andpharmaceutically acceptable salts thereof, wherein

[0054] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;

[0055] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain allyl; and

[0056] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, —OH, —C₁-C₆ alkyl, —NO₂, —OC₁-C₆alkyl, and —OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof; and

[0057] each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)— and where

[0058] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemnates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂, and where

[0059] n is an integer from 2 to 13, preferably 2 to 5.

[0060] In one embodiment of the invention, the compound of formula (IV)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0061] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (IV) selectedfrom the group consisting ofD-2-O-butyryl-1,2,4,5,6-pentahydrobenzoxirane 1,4,5,6-tetrakisphosphateoctakis-(acetoxymethyl) ester,D-2-O-butyryl-2,3,4,5,6-pentahydrobenzoxirane 3,4,5,6-tetra-kisphosphateoctakis(acetoxymethyl)ester; D-2-O-butyryl-1,2,4,5,6-pentahydrochroman1,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester;D-2-O-butyryl-2,3,4,5,6-pentahydrochroman 3,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester; or a racemate or a pharmaceuticallyacceptable salt thereof.

[0062] The present invention still further provides novel compounds,useful for regulating chloride secretion and/or inhibiting inflammation,having the general formula (V):

[0063] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain allyl and —OC₁-C₂₀ straight or branched chainalkyl, —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂;

[0064] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0065] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, —OH, C₁-C₆ alkyl, —NO₂, —OC₁-C₆alkyl, and —OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof; and

[0066] each V is independently selected from a group consisting of —C(X—C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)—; and

[0067] m is an integer from 2 to 15, preferably from 2 to 5; and

[0068] each W is independently selected from a group consisting of—C(X)—, —C(X)—C(X)(Y)—, or —C(X)—C(X)(Y)—C(X)(Y)—; and

[0069] n is an integer from 1 to 15, preferably from 1 to 3; and

[0070] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂.

[0071] The present invention further provides compositions, useful forregulating chloride secretion and/or inhibiting inflammation, comprisinga therapeutically effective amount of compounds of formula (V) or aracemate thereof, and pharmaceutically acceptable salts thereof. Suchcompositions can also comprise a pharmaceutically acceptable carrier orvehicle.

[0072] The invention further provides methods for regulating chloridesecretion and/or inhibiting inflammation in a cell or tissue with aneffective amount of a compound of formula (V), or regulating chloridesecretion and/or inhibiting inflammation in a human or animal patient inneed of such increased chloride secretion using a therapeuticallyeffective amount of a compound of formula (V), and racemates,stereoisomers and pharmaceutically acceptable salts thereof.

[0073] In one embodiment of the invention, the compound of formula (V)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0074] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (V) selected fromthe group consisting of (1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3;4,5-tetrakis[[is(acetoxymethoxy)phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.0^(1,6)]-dodecane;or a racemate or a pharmaceutically acceptable salt thereof.

[0075] The compounds of formulas (I), (II), (III), (IV) and (V) may beadministered by a variety of methods including orally, sublingually,intranasally, intramuscularly, intravenously, subcutaneously,intravaginally, transdermally, rectally, by inhalation, or as amouthwash.

[0076] The present invention may be understood more fully by referenceto the following figures, detailed description and illustrative exampleswhich are intended to exemplify non-limiting embodiments of theinvention.

DESCRIPTION OF THE DRAWINGS

[0077]FIG. 1 is a reaction scheme showing a representative route for thesynthesis of 2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(proprioxymethyl)ester (also referred to herein as “TMX”), asdescribed in Example 1.

[0078]FIG. 2 is a reaction scheme showing a representative route for thesynthesis of rac-(3aS, 4R, 5S, 6R, 7S,7aS)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol(also referred to herein as “Compound B”), as described in Example 2.

[0079]FIG. 3 is a reaction scheme showing a representative route for thesynthesis, of rac-(1S, 2R, 3S, 4R, 5S, 6S)4,5,6,7-tetrakis[[bis(acetoxyethoxy)-phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane (alsoreferred to herein as “Compound A”), as described in Example 3.

[0080]FIG. 4 is a reaction scheme showing a representative route for thesynthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxymethoxy)-phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane(also referred to herein as “Compound D”), as described in Example 4.

[0081]FIG. 5 is a reaction scheme showing a representative route for thesynthesis of(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(also referred to herein as “Compound C”), as described in Example 5.

[0082]FIG. 6 is a reaction scheme showing a representative route for thesynthesis of(4aα,6β,7α,8β,8aα)-5,6,7,8-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]4a-butyryloxy-octahydro-chromen(also referred to herein as “Compound G”), as described in Example 6.

[0083]FIG. 7 is a reaction scheme showing a representative route for thesynthesis of a tricyclo compound, (1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis-[[bis(acetoxymethoxy)phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.0^((1,6))]-dodecane,as described in Example 7.

[0084]FIG. 8 is a graph showing transepithelial current stimulation byinositol derivative composition Library B, as described in Example 8.Monolayers were plated and mounted in modified Ussing chambers.Indomethacin (10 μM) and amiloride (100 μM) were added to the apicalcompartment and the baseline allowed to stabilize for 10-20 minutesprior to addition of the test composition. Final concentration ofDMSO/DMSO-5% pluronic F-127=0.12%). Amiloride caused a decrease inI_(sc) in 19 of 23 monolayers of 1.01 μAmps/cm².

[0085]FIG. 9A is a bar graph showing the increase in short circuitcurrent (I_(sc)) in monolayers of T₈₄ cells in Ussing chambers exposedto differing concentrations of D-2,3,5-tri-O-butyryl-myo-inositol1,4,6-trisphosphate hexakis(proprionoxymethyl) ester (“TMX”; ent-TMX isthe enantiomer of TMX), as described in Example 9. FIG. 9B is a graphshowing that TMX induces a prolonged increase in I_(sc) similar tocyclic AMP in colonic epithelial cell line T₈₄. FIG. 9C is a graphshowing the effect of acute addition of TMX in human CF nasal epithelia.

[0086]FIG. 10A is a graph showing the effect ofrac-2-O-butyryl-2-C-propyl-1-O-propyl-myo-inositol3,4,5,6-tetrakisphosphate-octakis(acetoxymethyl)ester (“Compound A”) onthe stimulation of an increase I_(sc) in CFHNE (passage 3) in comparisonto response to ATP, as described in Example 8. Example 10B is a graphshowing prolonged increase in I_(sc) following ATP addition in CFHNE.

[0087]FIG. 11 is a graph showing that TMX and Compound B reverse thenegative component of the ATP effect on I_(sc) in CFHNE. Data are theaverage of duplicates. The solid line indicates preincubation withCompound B at 400 μM.

[0088]FIG. 12 is a graph showing that Compound B triggers an increase inI_(sc) in T₈₄ cells, as described in Example 9. Compound B was appliedacutely to the apical surface of T₈₄ monolayers mounted in Ussingchambers at the indicated time, as described in Example 9. After 25 min.10⁻⁴ M carbachol was added. Data plotted as ΔI_(sc) (μA/m²) representsone monolayer.

[0089]FIG. 13 is a graph showing monolayers of colonic epithelia (T₈₄)preincubated for 30 minutes with Compound B have a prolonged increase inI_(sc) response to the calcium elevating agents, carbachol andthapsigargin, as described in Example 9.

[0090]FIG. 14 is a graph showing that preincubation of colonicepithelia, T₈₄, with 200 μM Compound C for 30 minutes reduces themagnitude of the subsequent increase in Isc in response to carbachol, asdescribed in Example 9.

[0091]FIG. 15 is a graph showing that preincubation of colonicepithelia, T₈₄, with 400 μM Compound C for 30 minutes reduces themagnitude of the subsequent increase in I_(sc) in response to carbacholbut partially restores the response of the monolayers to thapsigargin,as described in Example 9. Carbachol has been shown to reduce thesubsequent I_(sc) response of T₈₄ monolayers to thapsigargin.

[0092]FIG. 16 is a graph showing that preincubation of T₈₄ monolayerswith Compound A (400 μM) partially restores the I_(sc) response tothapsigargin following carbachol, as described in Example 9.

[0093]FIG. 17 is a graph showing that Compound D has a small acuteeffect increasing basal I_(sc) in colonic epithelia, T₈₄, as describedin Example 9. However, there is little effect on subsequentcarbachol-stimulated I_(sc).

[0094]FIG. 18 is a graph showing that preincubation with Compound Greverses the carbachol-mediated inhibition of thapsigargin-stimulatedincrease in I_(sc) in T₈₄ colonic epithelia, as described in Example 9.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0095] In accordance with the present invention, novel compounds andmethods useful for regulating chloride secretion and/or inhibitinginflammation of epithelial cells are provided. In one aspect of theinvention, the novel compounds have the general formula (I):

[0096] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, and —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0097] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0098] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0099] The present invention further provides compositions, useful formodulating chloride secretion, comprising a therapeutically effectiveamount of compounds of formula (I) or a racemate thereof, andpharmaceutically acceptable salts thereof. As used herein, the terms“modulating chloride secretion” and “regulating chloride secretion” areused in relationship to compounds of the invention that enhance oractivate chloride secretion from treated cells or tissues, as well ascompounds that inhibit chloride secretion from treated cells or tissues.The actual in vivo activity of a compound of the invention in themodulation of chloride secretion may be reliably predicted by thechloride secretion assays disclosed herein, such as the assays ofExamples 8 and 9, as well as other chloride secretion assays well knownto those skilled in the art. In one aspect of the present invention,presently preferred compounds of the invention represented by formulas(I)-(V) are effective for activating or enhancing chloride secretion incells or tissues treated with the compounds.

[0100] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0101] The invention further provides methods for regulating chloridesecretion, comprising administering to a patient in need of suchincreased or decreased chloride secretion a therapeutically effectiveamount of a compound of formula (I), or a racemate thereof, or apharmaceutically acceptable salt thereof, wherein:

[0102] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0103] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0104] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0105] In one embodiment of the invention, the compound of formula (I)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0106] The invention further provides methods for regulating chloridesecretion in a cell or tissue with an effective amount of a compound offormula (I) or a racemate thereof, or a pharmaceutically acceptable saltthereof, wherein:

[0107] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0108] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0109] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0110] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (I) selected fromthe group consisting of D-2,3,5-tri-O-butyryl-myo-inositol1,4,6-trisphosphate hexakis(acetoxymethyl)ester;D-2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(propionoxymethyl) ester, or a racemate thereof, or apharmaceutically acceptable salt thereof.

[0111] In one embodiment of the invention, the compound of formula (I)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0112] The present invention still further provides novel compounds,useful for stimulating chloride secretion of epithelial cells areprovided, having the general formula (II):

[0113] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, and —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0114] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0115] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof with the proviso that the compound of formula (II)is not D-3,6-di-O-butyryl-myo-inositol 1,2,4,5-tetrakisphosphateoctakis(acetoxymethyl) ester, D-1,4-di-O-butyryl-myo-inositol2,3,5,6-tetrakisphosphate octakis(acetoxymethyl)ester or a racematethereof.

[0116] The present invention further provides compositions, useful forregulating chloride secretion, comprising a therapeutically effectiveamount of compounds of formula (II) or a racemate thereof, andpharmaceutically acceptable salts thereof, with the proviso that thecompound of formula (II) is not D-3,6-di-O-butyryl-myo-inositol1,2,4,5-tetrakisphosphate octakis(acetoxymethyl) ester;D-1,4-di-O-butyryl-myo-inositol 2,3,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester or a racemate thereof.

[0117] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0118] The invention further provides methods for regulating chloridesecretion, comprising administering to, a patient in need of suchmodified chloride secretion a therapeutically effective amount of acompound of formula (II), or a racemate thereof, or a pharmaceuticallyacceptable salt thereof, wherein:

[0119] each R₁ is independently selected from the groups consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0120] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0121] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0122] In one embodiment of the invention, the compound of formula (II)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0123] The invention further provides methods for regulating chloridesecretion in a cell or tissue with an effective amount of a compound offormula (II) or a racemate thereof, or a pharmaceutically acceptablesalt thereof, wherein:

[0124] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl;

[0125] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0126] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof

[0127] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (II) selectedfrom the group consisting of D-3,6-di-O-butyryl-myo-inositol1,2,4,5-tetrakisphosphate octakis(propion-oxymethyl) ester;D-1,4-di-O-butyryl-myo-inositol 2,3,5,6-tetrakisphosphateoctakis-(propionoxymethyl) ester or a racemate or a pharmaceuticallyacceptable salt thereof.

[0128] In one embodiment of the invention, the compound of formula (II)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0129] The present invention still further provides novel compounds,useful for stimulating chloride secretion of epithelial cells areprovided, having the general formula (III):

[0130] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂; or each R₁ may be taken together to form a cyclic structure;

[0131] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₄)R₄)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0132] each R₃ is independently selected from a group consisting of—C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight or branchedchain alkenyl or alkynyl, —C₁-C₁₁—X—Y, where each X is independentlyselected from a group consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—,—CO(S)—, —OC(O)—, —SC(O)—, OC(S)—, or —NH—, and where Y is independentlyselected from a group consisting of —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof

[0133] each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0134] The present invention further provides compositions, useful forregulating chloride secretion, comprising a therapeutically effectiveamount of compounds of formula (III) or a racemate thereof, andpharmaceutically acceptable salts thereof.

[0135] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0136] The invention further provides methods for regulating chloridesecretion, comprising administering to a patient in need of suchmodified chloride secretion a therapeutically effective amount of acompound of formula (III), or a racemate thereof, or a pharmaceuticallyacceptable salt thereof, wherein

[0137] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;or each R₁ may be taken together to form a cyclic structure;

[0138] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₄)(₄)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0139] each R₃ is independently selected from a group consisting of—C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight or branchedchain alkenyl or alkynyl, —C₁-C₁₁—X—Y, where each X is independentlyselected from a group consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—,—CO(S)—, —OC(O)—, —SC(O)—, OC(S)—, or —NH—, and where Y is independentlyselected from a group consisting of —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof.

[0140] each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0141] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0142] The invention further provides methods for regulating chloridesecretion, comprising administering to a patient in need of suchmodulated chloride secretion a therapeutically effective amount of acompound of formula (III), or a racemate thereof, or a pharmaceuticallyacceptable salt thereof, wherein

[0143] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;or each R₁ may be taken together to form a cyclic structure;

[0144] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0145] each R₃ is independently selected from a group consisting of—C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight or branchedchain alkenyl or alkynyl, —C₁-C₁₁—X—Y, where each X is independentlyselected from a group consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—,—CO(S)—, —OC(O)—, —SC(O)—, OC(S)—, or —NH—, and where Y is independentlyselected from a group consisting of —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof.

[0146] each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0147] In one embodiment of the invention, the compound of formula (III)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0148] The invention further provides methods for regulating chloridesecretion in a cell or tissue with an effective amount of a compound offormula (III) or a racemate thereof, or a pharmaceutically acceptablesalt thereof wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, —OC₂-C₂₀ straight- or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂; or each R₁ may be taken together to form a cyclic structure;

[0149] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain allyl; and

[0150] each R₃ is independently selected from a group consisting of—C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight or branchedchain alkenyl or alkynyl, —C₁-C₁₁—X—Y, where each X is independentlyselected from a group consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—,—CO(S)—, —OC(O)—, —SC(O)—, OC(S)—, or —NH—, and where Y is independentlyselected from a group consisting of —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof.

[0151] each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R⁴ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R⁴, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof.

[0152] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (III) selectedfrom the group consisting ofD-2-O-butyryl-2-C-propyl-3-O-propyl-myo-inositol1,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester;D-2-O-butyryl-2-C-propyl-1-O-propyl-myo-inositol3,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester,D-2,3-O-isoproylidene-2-C-propyl-myo-inositol. 1,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester;D-1,2-O-isoproylidene-2-C-propyl-myo-inositol 3,4,5,6-tetrakisphosphateoctakis-(acetoxymethyl) ester; or a racemate or a pharmaceuticallyacceptable salt thereof.

[0153] The present invention still further provides novel compounds,useful for stimulating chloride secretion of epithelial cells areprovided, having the general formula (IV):

[0154] and racemates pharmaceutically acceptable salts thereof, whereineach. R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched, chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂;

[0155] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0156] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆′ alkyl groups, including racemates anddiastereomers thereof; and

[0157] each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)— and where

[0158] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂, and where

[0159] n is an integer from 2 to 13, preferably 2 to 5.

[0160] The present invention further provides compositions, useful forregulating chloride secretion, comprising a therapeutically effectiveamount of compounds of formula (I) or a racemate thereof, andpharmaceutically acceptable salts thereof. Such compositions can alsocomprise a pharmaceutically acceptable carrier or vehicle. The inventionfurther provides methods for regulating chloride secretion, comprisingadministering to a patient in need of such modulated chloride secretiona therapeutically effective amount of a compound of formula (IV), andracemates, stereoisomers and pharmaceutically acceptable salts thereof,wherein

[0161] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straighterbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;

[0162] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0163] each R₃ is independently selected from a group consisting ofhydrogen and. —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof; and

[0164] each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)— and where

[0165] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂, and where

[0166] n is an integer from 2 to 13, preferably 2 to 5.

[0167] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0168] The invention further provides methods for regulating chloridesecretion, comprising administering to a patient in need of suchmodified chloride secretion a therapeutically effective amount of acompound of formula (IV), and racemates, stereoisomers andpharmaceutically acceptable salts thereof, wherein

[0169] each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₁₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;

[0170] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0171] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof; and

[0172] each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)— and where

[0173] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂, and where

[0174] n is an integer from 2 to 13, preferably 2 to 5.

[0175] In one embodiment of the invention, the compound of formula (IV)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0176] The invention further provides methods for regulating chloridesecretion in a cell or tissue with an effective amount of a compound offormula (IV) or a racemate thereof, or a pharmaceutically acceptablesalt thereof, wherein

[0177] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainalkyl, —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂;

[0178] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0179] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups, including racemates anddiastereomers thereof; and

[0180] each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)Y)—, or —C(X)(Y)—C(X)(Y)—CM)(Y)— and where

[0181] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)allyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂, and where

[0182] n is an integer from 2 to 13, preferably 2 to 5.

[0183] In a preferred embodiment, the present methods comprisecontacting a cell or tissue with a compound of formula (IV) selectedfrom the group consisting ofD-2-O-butyryl-1,2,4,5,6-pentahydrobenzoxirane 1,4,5,6-tetrakisphosphateoctakis-(acetoxymethyl) ester,D-2-O-butyryl-2,3,4,5,6-pentahydrobenzoxirane 3,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester; D-2-O-butyryl-1,2,4,5,6-pentahydrochroman1,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester;D-2-O-butyryl2,3,4,5,6-pentahydrochroman 3,4,5,6-tetrakisphosphateoctakis-(acetoxymethyl) ester; or a racemate thereof, or apharmaceutically acceptable salt thereof.

[0184] The present invention still further provides novel compounds,useful for stimulating chloride secretion of epithelial cells areprovided, having the general formula (V):

[0185] and racemates, stereoisomers and pharmaceutically acceptablesalts thereof, wherein each R₁ is independently selected from the groupconsisting of hydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl,—C₂-C₂₀ straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀straight or branched chain alkyl and —OC₁-C₂₀ straight or branched chainallyl, —OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and—OPO(OR₂)₂;

[0186] each R₂ is independently selected from a group consisting ofhydrogen and —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and

[0187] each R₃ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₃, except hydrogen, being unsubstituted orsubstituted with one or more halogen, —OH, C₁-C₆ allyl, —NO₂, —OC₁-C₆alkyl, and —OC(O)C₁-C₆ allyl groups, including racemates anddiastereomers thereof; and

[0188] each V is independently selected from a group consisting of—C(X)(Y)—, —C(X(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)—; and

[0189] m is an integer from 2 to 15, preferably from 2 to 5; and

[0190] each W is independently selected from a group consisting of—C(X)—, —C(X)—C(X)(Y)—, or —C(X)—C(X)(Y)—C(X)(Y)—; and

[0191] n is an integer from 1 to 15, preferably from 1 to 3; and

[0192] X and Y are independently selected from a group consisting ofhydrogen, —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight orbranched chain alkenyl or alkynyl, or unsubstituted and substitutedbenzyl, and phenyl, including racemates and diastereomers thereof andhydrogen, —OH, —Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl,—CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or—CONHalkyl, or —CON(alkyl)(alkyl), —NH₂.

[0193] The present invention further provides compositions, useful forregulating chloride secretion and/or inhibiting inflammation, comprisinga therapeutically effective amount of compounds of formula (V) or aracemate thereof, and pharmaceutically acceptable salts thereof.

[0194] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0195] The invention further provides methods for regulating chloridesecretion, comprising administering to, a patient in need of suchmodified chloride secretion a therapeutically effective amount of acompound of formula (V), and racemates, stereoisomers andpharmaceutically acceptable salts thereof.

[0196] Such compositions can also comprise a pharmaceutically acceptablecarrier or vehicle.

[0197] The invention further provides methods for regulating chloridesecretion, comprising administering to a patient in need of suchmodified chloride secretion a therapeutically effective amount of acompound of formula (V), and racemates, stereoisomers andpharmaceutically acceptable salts thereof.

[0198] In one embodiment of the invention, the compound of formula (IV)is administered in a composition comprising a pharmaceuticallyacceptable carrier or vehicle. Such carrier or vehicle can include aliposomal formulation or a formulation in an aerosol.

[0199] The invention further provides methods for regulating chloridesecretion in a cell or tissue with an effective amount of a compound offormula (V) or a racemate thereof, or a pharmaceutically acceptable saltthereof.

[0200] As used herein, the term “alkyl” as used herein refers tobranched or straight chain alkyl groups comprising one to twenty carbonatoms (C₁-C₂₀) that are unsubstituted or substituted, e.g., with one ormore halogen groups, including, e.g., methyl, ethyl, propyl, isopropyl,n-butyl, t-butyl, neopentyl, trifluoromethyl, pentafluoroethyl and thelike.

[0201] The term “alkoxy” as used herein refers to RO— wherein R is alkylas defined above. Representative examples of alkoxy groups includemethoxy, ethoxy, t-butoxy, trifluoromethoxy and the like.

[0202] The term “alkenyl” as used herein refers to a branched orstraight chain groups comprising two to twenty carbon atoms which alsocomprise one or more carbon-carbon double bonds. Representative alkenylgroups include 2-propenyl (i.e., allyl), 3-methyl-2-butenyl,3,7-dimethyl-2,6-octadienyl, 4,8-dimethyl3,7-nonadienyl,3,7,11-trimethyl-2,6,10-dodecatrienyl and the like.

[0203] The term “alkynyl” as used herein refers to a branched orstraight chain comprising two to twenty carbon atoms that also comprisesone or more carbon-carbon triple bonds. Representative alkynyl groupsinclude ethynyl, 2-propynyl (propargyl), 1-propynyl and the like.

[0204] The term “halogen” or “halo” as used herein refers to iodo,bromo, chloro or fluoro.

[0205] The term “haloalkyl” as used herein refers to a lower alkylradical, as defined above, bearing at least one halogen substituent, forexample, chloromethyl, fluoro-ethyl or trifluoromethyl and the like.

[0206] In presently particularly preferred embodiments, theIns(3,4,5,6)P₄ antagonists of the invention are designed to be deliveredintracellularly, such as by concealing the negatively charged phosphategroups with bioactivatable esters, such as acetoxymethylesters(AM-esters), and the hydroxy groups with alkyl groups, such asbutyrates, where necessary. These masking groups have previously beenshown to permit passive diffusion of the inositol polyphosphatecompounds across the plasma membrane to the interior of the cell whereesterases cleave them and liberate the biologically active inositolpolyphosphate inside the cell. (See M. Vajanaphanich et al., Nature371:711 (1994); Rudolf, M. T. et al., “2-Deoxy derivative is a partialagonist of the intracellular messenger inositol3,4,5,6-tetrakisphosphate in the epithelial cell line T84” J Med Chem41:363544 (1998)).

[0207] In general, the compounds of the invention can be prepared byprocesses generally known in the art, and by the processes illustratedin Schemes I (FIG. 1), II (FIG. 2), III (FIG. 3), IV (FIG. 4), V (FIG.5)and VI (FIG. 6). See, for example, T. Meyerdierks, H. H. Gillandt, andCarsten Schultz, A Membrane-Permeant, Bioactivatable Derivative OfIns(1,4,6)P₃ Elevates Intracellular Calcium Levels Of PC12 Cells,(1999); and C. Schultz et al., Membrane-Permeant, BioactivatableDerivatives of Inositol Polyphosphates and phosphoinositides, inPhosphoinositides: Chemistry, Biochemistry and Biomedical Applications,K. S. Bruzik, Ed. Am. Chem. Soc., Symp. Ser., 718, 232-243 (1998); forprocedures for the production of compounds of structures I and II. Inaddition, representative compounds of structures II-IV are described infurther detail in the figures and examples.

[0208] The compounds of the present invention can be used in the form ofsalts derived from inorganic or organic acids. These salts include butare not limited to the following: acetate, adipate, alginate, citrate,aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate,camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate,ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate,heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, nicotinate, 2-napthalenesulfonate, oxalate, pamoate,pectinate, persulfate, 3-phenylproionate, picrate, pivalate, propionate,succinate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate.Also, the basic nitrogen-containing groups can be quaternized with suchagents as loweralkyl halides, such as methyl, ethyl, propyl, and butylchloride, bromides, and iodides; dialkyl sulfates like dimethyl,diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl,lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkylhalides like benzyl and phenethyl bromides, and others. Water oroil-soluble or dispersible products are thereby obtained.

[0209] Examples of acids which may be employed to form pharmaceuticallyacceptable acid addition salts include such inorganic acids ashydrochloric acid, sulphuric acid and phosphoric acid and such organicacids as oxalic acid, maleic acid, succinic acid and citric acid. Basicaddition salts can be prepared in situ during the final isolation andpurification of the compounds of formulas (I)-(V), or separately byreacting carboxylic acid moieties with a suitable base such as thehydroxide, carbonate or bicarbonate of a pharmaceutical acceptable metalcation or with ammonia, or an organic primary, secondary or tertiaryamine. Pharmaceutical acceptable salts include, but are not limited to,cations based on the alkali and alkaline earth metals, such as sodium,lithium, potassium, calcium, magnesium, aluminum salts and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. Other representative organicamines useful for the formation of base addition salts includediethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazineand the like.

[0210] The compounds of the invention are useful in vitro for modulatingchloride secretion in a cell or tissue, and in vivo in human and animalhosts for obtaining increases and decreases in chloride secretion. Thecompounds may be used alone or in compositions together with apharmaceutically acceptable carrier.

[0211] Thus, in one aspect, the present invention provides methods oftreatment of cystic fibrosis in a subject in need of such treatment byadministering an inositol polyphosphate as given above to said subjectin an amount effective to hydrate lung mucus secretions. In anotheraspect, the present invention provides methods of treating chronicbronchitis in a subject in need of such treatment by administering aninositol polyphosphate as given above to said subject in an amounteffective to hydrate lung mucus secretions. In another aspect, thepresent invention provides methods of treating asthma in a subject inneed of such treatment by administering an inositol polyphosphate analogas given above to said subject in an amount effective to hydrate lungmucus secretions. In another aspect, the present invention providesmethods of combating chronic obstructive pulmonary disorder by inositolpolyphosphate analog as given above to said subject in an amounteffective to hydrate lung mucus secretions. In another aspect, thepresent invention provides methods of combating sinusitis byadministering an inositol polyphosphate analog as given above to saidsubject in an amount effective to hydrate sinus mucus secretions. Inanother aspect, the present invention provides methods of combatingdiarrhea by administering an inositol polyphosphate analog as givenabove to said subject in an amount effective to reduce intestinal fluidsecretion. In another aspect, the present invention provides methods ofimproving mucosal hydration in mucous membranes where insufficienthydration can lead to discomfort, such as in dryness of the eyes ordryness of the vaginal mucosa. In another aspect, the present inventionprovides methods of combating swelling by administering an inositolpolyphosphate analog as given above to said subject in an amounteffective to modulate fluid movement across membranes. In anotheraspect, the present invention provides methods of combating inflammationsuch as to reduce inflammation in such chronic conditions as asthma,adult respiratory distress syndrome, infant respiratory distresssyndrome, osteoarthritis, rheumatoid arthritis, inflammatory boweldisease, psoriasis, dermatitis, other inflammatory conditions of theskin. In another aspect, the present invention provides methods ofaccelerating wound healing, by combating inflammation and swelling. Inyet other aspects, the present invention provides methods of use of theactive compounds for the treatment of chronic conditions shown to bealleviated by anti-inflammatory treatment such as arterioschlerosis andAlzheimers Disease. In yet other aspects the present invention providesmethods of regulating fluid flux across membranes of the kidney. In yetother aspects, the present invention provides methods of use of anactive compound as disclosed herein for the manufacture of a medicamentfor the prophylactic or therapeutic treatment of cystic fibrosis in asubject in need of such treatment. In yet other aspects, the presentinvention provides methods of use of an the active compounds asdisclosed herein for the manufacture of a medicament for theprophylactic or therapeutic treatment of chronic bronchitis or thereduction of inflammation in a subject in need of such treatment. In yetother aspects, the present invention provides methods of use of an theactive compounds as disclosed herein for the manufacture of a medicamentfor the prophylactic or therapeutic treatment of asthma in a subject inneed of such treatment.

[0212] When administered to a patient, e.g., a mammal for veterinary useor to a human for clinical use, the inositol derivatives are preferablyadministered in isolated form. By “isolated” is meant that prior toformulation in a composition, the inositol derivatives are separatedfrom other components of either (a) a natural source such as a plant orcell culture, or (b) a synthetic organic chemical reaction mixture.Preferably, via conventional techniques, the inositol derivatives arepurified.

[0213] When administered to a patient, e.g., a mammal for veterinary useor to a human for clinical use, or when made to contact a cell ortissue, the inositol derivatives can be used alone or in combinationwith any physiologically acceptable carrier or vehicle suitable forenteral or parenteral delivery. Where used for enteral, parenteral,topical, otic, ophthalmologic, intranasal, oral, sublingual,intramuscular, intravenous, subcutaneous, intravaginal, transdermal, orrectal administration, the physiologically acceptable carrier or vehicleshould be sterile and suitable for in vivo use in a human, or for use ina veterinary clinical situation.

[0214] In addition, the inositol derivatives can be administered topatients or contacted with a cell or tissue in liposome formulations,which facilitate their passage through cell membranes. Accordingly, therelative impermeability of cell membranes to relatively polar inositolderivatives can be overcome by their encapsulation in liposomalformulations. The characteristics of liposomes can be manipulated bymethods known to those of ordinary skill in the art, such that size,membrane fluidity, tissue targeting, and compound release kinetics areadapted to the particular condition (Georgiadis, NIPS 4:146 (1989)).Liposomes of various sizes and compositions that encapsulate theinositol derivatives for delivery can be achieved by methods known tothose skilled in the art (See, for example, Hope et al., Biochem.Biophys. Acta 812:55 (1985); Hernandez, et al., J. Microencapsul. 4:315(1987); Singh, et al., Cancer Lett. 84:15 (1994); and Dipali, et al., J.Pharm. Pharmacol. 48:1112 (1996)).

[0215] The inositol derivatives can be used in the form of apharmaceutical preparation, for example, in solid, semisolid or liquidform, that contains at least one of the inositol derivatives of thepresent invention as a bioactive component, alone or in combination withan anti-inflammatory compound, in admixture with a carrier, vehicle oran excipient suitable for enteral or parental administration. Suchanti-inflammatory compounds useful in this regard include, but are notlimited to, non-steroidal anti-inflammatory drugs such as salicylicacid, acetylsalicylic acid, methyl salicylate, diflunisal, salsalate,olsalazine, sulfasalazine, acetaminophen, indomethacin, sulindac,etodolac, mefenamic acid, meclofenamate sodium, tolmetin, ketorolac,dichlofenac, ibuprofen, naproxen, naproxen sodium, fenoprofen,ketoprofen, flurbinprofen, oxaprozin, piroxicam, meloxicam, ampiroxicam,droxicam, pivoxicam, tenoxicam, nabumetome; phenylbutazone,oxyphenbutazone, antipyrine, aminopyrine, apazone and nimesulide;leukotriene antagonists including, but not limited to, zileuton,aurothioglucose, gold sodium thiomalate and auranofin; and otheranti-inflammatory agents including, but not limited to, colchicine,allopurinol, probenecid, sulfinpyrazone and benzbromarone.

[0216] In addition, the inositol derivatives of the present inventionmay be compounded, for example with a pharmaceutically acceptablecarrier or vehicle for solid compositions such as tablets, pellets orcapsules; capsules containing liquids; suppositories; solutions;emulsions; aerosols; sprays; suspensions or any other form suitable foruse. Suitable carriers and vehicles include, for example, sterile water,sterile physiological saline, gum acacia, gelatin, starch paste, talc,keratin, colloidal silica, urea and the like. In addition, auxiliary,stabilizing, thickening, lubricating and coloring agents may be used.The inositol derivatives are present in the compositions in atherapeutically effective amount, i.e., an amount sufficient to restorenormal mucosal secretions.

[0217] The compositions of this invention may be administered by avariety of methods including orally, sublingually, intranasally,intramuscularly, intravenously, subcutaneously, intravaginally,transdermally, rectally, by inhalation, or as a mouthwash in dosage unitformulations containing conventional nontoxic pharmaceuticallyacceptable carriers, adjuvants, and vehicles as desired. Topicaladministration may also involve the use of transdermal administrationsuch as transdermal patches or ionophoresis devices. The preferred modeof administration is left to the discretion of the practitioner, andwill depend in-part upon the desired site of action.

[0218] For example, when the cystic fibrosis, chronic bronchitis orasthma affects the function of the lungs, the inositol derivatives canbe administered as an atomized aerosol, via a nebulizer, or viaperfusion in a fluorocarbon or synthetic pulmonary surfactant;alternatively, the inositol derivatives can be administeredintravenously directly. Thus, the active compounds disclosed herein maybe administered to the lungs of a patient by any suitable means, but arepreferably administered by generating an aerosol comprised of respirableparticles, the respirable particles comprised of the active compound,which particles the subject inhales. The respirable particles may beliquid or solid. The particles may optionally contain other therapeuticingredients such as a sodium channel blocker as noted above, with thesodium channel blocker included in an amount effective to inhibit thereabsorption of water from airway mucous secretions. The particles mayoptionally contain other therapeutic ingredients such as lantibiotics asdescribed in U.S. Pat. Nos. 5,512,269 and 5,716,931 or UridineTriphosphate Analogs as described in U.S. Pat. No. 5,292,498, nitricoxide inhibitors as described in U.S. Pat. No. 5,859,058, dinucleotidesas described in U.S. Pat. No. 5,935,555, or organic acids as describedin U.S. Pat. No. 5,908,611. Particles comprised of active compound forpracticing the present invention should include particles of respirablesize: that is, particles of a size sufficiently small to pass throughthe mouth and larynx upon inhalation and into the bronchi and alveoli ofthe lungs. In general, particles ranging from about 0.5 to 10 microns insize (more particularly, less than about 5 microns in size) arerespirable. Particles of non-respirable size which are included in theaerosol tend to deposit in the throat and be, swallowed, and thequantity of non-respirable particles in the aerosol is preferablyminimized. For nasal administration, a particle size in the range of10-500 μm is preferred to ensure retention in the nasal cavity.

[0219] Liquid pharmaceutical compositions of active compound forproducing an aerosol can be prepared by combining the active compoundwith a suitable vehicle, such as sterile pyrogen free water. Othertherapeutic compounds, such as a sodium channel blocker, may optionallybe included. Solid particulate compositions containing respirable dryparticles of micronized active compound may be prepared by grinding dryactive compound with a mortar and pestle, and then passing themicronized composition through a 400 mesh screen to break up or separateout large agglomerates. A solid particulate composition comprised of theactive compound may optionally contain a dispersant that serves tofacilitate the formation of an aerosol. A suitable dispersant islactose, which may be blended with the active compound in any suitableratio (e.g., a 1 to 1 ratio by weight). Again, other therapeuticcompounds, such as amiloride, may also be included.

[0220] The dosage of active compound for prophylaxis or treatment oflung disease will vary depending on the condition being treated and thestate of the subject, but generally may be an amount sufficient toachieve dissolved concentrations of active compound on the airwaysurfaces of the subject of from about 10⁻⁹ to 10⁻³ Moles/liter, and morepreferably from 10⁻⁷ to 10⁻⁵ Moles/liter. Depending on the solubility ofthe particular formulation of active compound administered, the dailydose may be divided among one or several unit dose administrations.Preferably, the daily dose is a single unit dose, which is preferablyadministered from 1 to 3 times a week. Treatments may continue week toweek on a chronic basis as necessary (i.e., the active agent can beadministered chronically). Administration of the active compounds may becarried out therapeutically (i.e., as a rescue treatment) orprophylactically, but preferably the compounds are administeredprophylactically, either before substantial lung blockage due toretained mucus secretions has occurred, or at a time when such retainedsecretions have been at least in part removed, as discussed above.

[0221] Aerosols of liquid particles comprising the active compound maybe produced by any suitable means, such as with a nebulizer. See, e.g.,U.S. Pat. No. 4,501,729. Nebulizers are commercially available devicesthat transform solutions or suspensions of the active ingredient into atherapeutic aerosol mist either by means of acceleration of a compressedgas; typically air or oxygen, through a narrow venturi orifice or bymeans of ultrasonic agitation. Suitable formulations for use innebulizers consist of the active ingredient in a liquid carrier, theactive ingredient comprising up to 40% w/w of the formulation, butpreferably less than 20% w/w. the carrier is typically water or a diluteaqueous alcoholic solution, preferably made isotonic with body fluids bythe addition of, for example, sodium chloride. Optional additivesinclude preservatives if the formulation is not prepared sterile, forexample, methyl hydroxybenzoate, antioxidants, flavoring agents,volatile oils, buffering agents and surfactants. Aerosols of solidparticles comprising the active compound may likewise be produced withany solid particulate medicament aerosol generator. Aerosol generatorsfor administering solid particulate medicaments to a subject produceparticles that are respirable, as explained above, and generate a volumeof aerosol containing a predetermined metered dose of a medicament at arate suitable for human administration. One illustrative type of solidparticulate aerosol generator is an insufflator. Suitable formulationsfor administration by insufflation include finely comminuted powdersthat may be delivered by means of an insufflator or taken into the nasalcavity in the manner of a snuff. In the insufflator, the powder (e.g., ametered dose thereof effective to carry out the treatments describedherein) is contained in capsules or cartridges, typically made ofgelatin or plastic, which are either pierced or opened in situ and thepowder delivered by air drawn through the device upon inhalation or bymeans of a manually-operated pump. The powder employed in theinsufflator consists either solely of the active ingredient or of apowder blend comprising the active ingredient, a suitable powderdiluent, such as lactose, and an optional surfactant. The activeingredient typically comprises from 0.1 to 100 w/w of the formulation. Asecond type of illustrative aerosol generator comprises a metered doseinhaler. Metered dose inhalers are pressurized aerosol dispensers,typically containing a suspension or solution formulation of the activeingredient in a liquified propellant. During use these devices dischargethe formulation through a valve adapted to deliver a metered volume,typically from 10 to 150 μl, to produce a fine particle spray containingthe active ingredient. Suitable propellants include certainchlorofluorocarbon compounds, for example, dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof.The formulation may additionally contain one or more co-solvents, forexample, ethanol, surfactants, such as oleic acid or sorbitan trioleate,antioxidants and suitable flavoring agents. The aerosol, whether formedfrom solid or liquid particles, may be produced by the aerosol generatorat a rate of from about 10 to 150 liters per minute, more preferablyfrom about 30 to 150 liters per minute, and most preferably about 60liters per minute. Aerosols containing greater amounts of medicament maybe administered more rapidly.

[0222] Where the condition of the subject to be treated affects thegastrointestinal tract, the inositol derivatives can be administeredrectally via enema or suppository, or orally in the form of a tablet orcapsule formulated to prevent dissolution prior to entry into theafflicted portion of the gastrointestinal tract; when the cysticfibrosis affects vaginal secretions, the inositol derivatives can beadministered intravaginally, in the form of a douche.

[0223] Compositions for oral delivery may be in the form of tablets,pills, troches, lozenges, aqueous or oily suspensions, granules orpowders, emulsions, capsules, syrups or elixirs. Orally administeredcompositions may contain one or more agents, for example, sweeteningagents such as fructose, aspartame or saccharin; flavoring agents suchas peppermint, oil of wintergreen, or cherry; coloring agents; andpreserving agents, to provide a pharmaceutically palatable preparation.Moreover, compositions in tablet form may be coated to delaydisintegration and absorption in the gastrointestinal tract therebyproviding a sustained action over an extended period of time.Selectively permeable membranes surrounding an osmotically activedriving compound are also suitable for orally administered compositions.In these later platforms, fluid from the environment surrounding thecapsule is imbibed by the driving compound, which swells to displace theagent or agent composition through an aperture. These delivery platformscan provide an essentially zero order delivery profile as opposed to thespiked profiles of immediate release formulations. A time delay materialsuch as glycerol monostearate or glycerol stearate may also be used.Liquid dosage forms for oral administration may include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions may also comprise adjuvants, such as wetting agents,emulsifying and suspending agents, and sweetening, flavoring, andperfuming agents.

[0224] Injectable preparations, for example, sterile injectable aqueousor oleagenous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a nontoxic parenterally acceptable diluent or solvent,for example, as a solution in ⅓-propanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solution,and isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordi-glycerides. In addition, fatty acids such as oleic acid find use inthe preparation of injectables.

[0225] Suppositories for rectal administration of the drug can beprepared by mixing the drug with a suitable nonirritating excipient suchas cocoa butter and polyethylene glycols, which are solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum and release the drug.

[0226] Aqueous suspensions containing the inositol derivatives may alsocontain one or more preservatives, such as, for example, ethyl orn-propyl-p-hydroxy-benzoate, one or more coloring agents, flavoringagents or sweetening agents.

[0227] Because the inositol derivatives are in the form oftetrakisphosphate, heptakis or octakis(acetoxymethyl or ethyl)esters,and because the inositol derivatives can contain —C₁-C₂₀ straight orbranched chain alkyl, —OC(O)C₁-C₂₀ straight or branched chain alkyl or—OC₁-C₂₀ straight or branched chain alkyl groups, the inositolderivatives possess enhanced lipophilic properties which allow forpassive diffusion across plasma membranes.

[0228] The compounds of the present invention can also be administeredin the form of liposomes. As is known in the art, liposomes aregenerally derived from phospholipids or other lipid substances.Liposomes are formed by mono- or multi-lamellar hydrated liquid crystalsthat are dispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additiona compound of the present invention, stabilizers, preservatives,excipients, and the like. The preferred lipids are the phospholipids andphosphatidyl cholines (lecithins), both natural and synthetic. Methodsto form liposomes are known in the art. See, for example, Prescott, Ed.,Methods in Cell Biology, Volume XIV, Academic Press, New York, N. W.(1976), p.33 et seq.

[0229] Without being bound by any particular theory, it is believed thatthe inositol derivatives function as “prodrugs” of a metabolized form ofthe inositol derivatives that are the actual pharmacological agentresponsible for the promotion of chloride secretion. Such prodrugs, byvirtue of their being more lipophilic than the actual pharmacologicalagents themselves, can more easily penetrate plasma membranes. Oncewithin a secretory cell, the prodrugs are converted, generallyenzymatically, to the active pharmacological agent. In addition, becausein vivo conversion of a prodrug to its active pharmacological formgenerally occurs over a period of time, rather than instantaneously, theuse of prodrugs offers the patient or subject the benefit of a sustainedrelease of the pharmacological agent, generally resulting in a longerduration of action.

[0230] In a further embodiment, the present invention contemplates theuse of an inositol derivative when delivered at a dose of about 0.001mg/kg to about 100 mg/kg body weight, preferably from about 0.01 toabout 10 mg/kg body weight. The inositol derivatives can be delivered upto several times per day, as needed. Treatment can be continued,indefinitely to normalize mucosal hydration or chloride secretion orreduce excessive mucosal viscosity.

[0231] The amount of active ingredient that may be combined with thecarrier materials to produce a single dosage form will vary dependingupon the host treated and the particular mode of administration.

[0232] It will be understood, however, that the specific dose level forany particular patient will depend upon a variety of factors includingthe activity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, rate of excretion, drug combination, and the severity ofthe particular disease undergoing therapy.

[0233] While the compounds of the invention can be administered as thesole active pharmaceutical agent, they can also be used in combinationwith one or more other agents used in the treatment of the symptoms ofcystic fibrosis, chronic bronchitis, asthma, inflammation and the like.For alleviating mucosal viscosity resulting from cystic fibrosis, acomposition of the present invention may be administered that comprisesan inositol derivative of the invention together with an agent usefulfor the treatment of inflammation-accompanying condition. For instance,for the treatment of cystic fibrosis, such an agent can be mucolytics(e.g., Pulmozyme® and Mucomyste®, purinergic receptor agonists such asuridine triphosphate (UTP), agents that suppress the cystic fibrosistransmembrane regulator (CFTR) premature stop mutation such asgentamycin, agents correcting the Delta F508 processing defect alsoknown as “protein assist therapies” such as CPX™ (SciClone),Phenylbutyrate (Ucyclyd Pharma), INS365, and genestein, and/or agentsfor the treatment of the accompanying infection such as tobramycin oraerosolized tobramycin (Tobi™), meropenem, RSV vaccine, IB605, Pa1806,anti-inflammatory agents such as DHA, rHEI, DMP777, IL10 (Tenovil)and/or agents triggering alternate chloride channels such aslantibiotics such as Duramycin (Moli901—Molichem Medicines), oromeprazole, and/or purinergic agonists such as nucleotide ordinucleotide analogs, or agents affecting sodium transport such asamiloride, and/or agents affecting pH such as organic acids.

[0234] For the treatment of asthma, such agents can becorticosteroids—such as fluticasone proprionate (Flovent®, FloventRotadisk®), budesonide (Pulmocort Turbuhaler®), flunisolide (Aerobid®),triamcinolone acetonide (Azmacort®), beclomethasone MDI (Beclovent®),Antileukotrienes such as Zafirlukast (Accolate®, Zeneca®), Zileuton(Zyflo®), Montelukast or other therapies such as Methotrexate,Troleandomycin, Gold, Cyclosporine, 5′ lipoxygenase inhibitors,bronchodilators, or immunotherapeutic agents.

[0235] CPX is a caffeine-like compound being investigated by SciClone.In laboratory studies it appears to increase chloride secretion in CFtissues that have the delta F508 mutation, but not in tissues with othermutations or normal epithelial cells. It is unknown whether it would beeffective in actual patients. Even if so, it would not benefit the 30%of CF sufferers who have other mutations.

[0236] Phenylbutyrate is a compound developed by Ucyclyd Pharma thattargets the protein generated by the delta F508 mutation. The CysticFibrosis Foundation is currently sponsoring a Phase I clinical trial ofthe drug at the Johns Hopkins University. However, because highconcentrations are necessary to be effective and the compound has anunappealing odor, other active analogs are currently being sought.

[0237] Duramycin is being developed by Molichem Medicines and, likephenylbutyrate, is reported to facilitate insertion of the delta F508CFTR into the plasma membrane. Unfortunately, duramycin is not effectivein the 30% of CF patients who have a genetic mutation other than deltaF508.

[0238] Purinergic (P2Y2) receptor agonists such as adenosinetriphosphate (ATP) and uridine triphosphate (UTP) stimulatecalcium-dependent chloride channels (not CFTR channels). They arecurrently being investigated by researchers at the University of NorthCarolina (under the auspices of Inspire Pharmaceuticals, Inc.) andindependently at Johns Hopkins University. Early trials indicate thatthis strategy could be useful in the treatment of cystic fibrosis andother chronic obstructive pulmonary disorders. However, theeffectiveness of this approach may be limited by inflammation-relatedinhibitory signals.

[0239] The compounds of the invention may also be administered incombination with one or more sodium channel blockers. Sodium channelblockers that may be used in the present invention are typicallypyrazine diuretics such as amiloride, as described in U.S. Pat. No.4,501,729. The term “amiloride” as used herein includes thepharmaceutically acceptable salts thereof, such as (but not limited to)amiloride hydrochloride, as well as the free base of amiloride. Thequantity of amiloride included may be an amount sufficient to achievedissolved concentrations of amiloride on the airway surfaces of thesubject of from about 10⁻⁷ to about 10⁻³ Moles/liter, and morepreferably from about 10⁻⁶ to about 10⁻⁴ Moles/liter.

[0240] The methods of the present invention may also further comprisethe step of removing retained mucus secretions from the lungs of thesubject prior to the step of administering the active agent. Thisfacilitates application of the active agent to the respiratory epitheliaduring the administering step. Such removal of retained mucus secretionscan be carried out by any suitable means, including postural drainage,antibiotic administration (e.g., intravenous or inhalationadministration of cephalosporin or aminoglycoside antibiotics such asTobramycin), and/or inhalation administration of DNase. In addition, thepresent invention may be carried out on patients such as children priorto decline of respiratory function (e.g., patients essentially free oflung blockage due to retained mucus secretions). Such patients can begenetically predisposed to becoming afflicted with lung disease (e.g.,cystic fibrosis) as hereinbefore described.

[0241] Alternatively, the compositions comprising an inositol derivativecan be administered in combination with, prior to, concurrent with orsubsequent to the administration of another agent useful for thetreatment of cystic fibrosis accompanying condition, as described above.

[0242] In addition, the inositol derivatives can be used for researchpurposes, for example, to investigate the mechanism and activity ofother agents thought to be useful for regulating mucosal hydration.

[0243] The foregoing may be better understood by reference to thefollowing examples, which are provided for, illustration and are notintended to limit the scope of the inventive concepts.

EXAMPLE 1 Synthesis of 2,3,5-Tri-O-butyryl-myo-inositol1,4,6-Trisphosphate Hexakis(proprioxymethyl) Ester (1). [TMX]

[0244] Referring to the reaction scheme shown in FIG. 1, the compound2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(proprioxymethyl)ester was synthesized as follows:

[0245] rac-1,4,6-Tri-O-benzyl-2,3-O-cyclohexylidene-myo-inositol(rac-4), rac-1,4,5-Tri-O-benzyl-2,3-O-cyclohexylidene-myo-inositol(rac-5) and rac-1,5-Di-O-benzyl-2,3-O-cyclohexylidene-myo-inositol

[0246] Under an argon atmosphere benzyl bromide (6 mL, 48 mmol) wasadded to a suspension of dry 1,2-O-cyclohexylidene-myo-inositol rac-3(2.08 g, 8 mmol), dry dibutyltin oxide (9.96 g, 40 mmol) and drytetrabutylammonium bromide (12.9 g, 40 mmol) in dry toluene (200 mL).The solution was stirred and heated under reflux in a Soxhlet apparatusfilled with activated molecular sieves (3 Å) for 24 h. After thereaction mixture was cooled to room temperature and Et₃N (14 mL, 96mmol) was added, the reaction was stirred for a further 20 h to destroythe excess of benzyl bromide. The solvent was evaporated, the residuewas dissolved in tert-butyl methyl ether (100 mL), and was washed twicewith sodium hydrogen carbonate (50 mL), with sodium hydrogen sulfate (50mL), with phosphate buffer (50 mL), and finally with brine (50 mL). Theorganic layer was dried over Na₂SO₄ and filtered. The solvent wasremoved under reduced pressure and the residual compounds were isolatedand purified by preparative HPLC [87% MeOH, 40 mL/min, t_(Ret)=16.54 min(rac-6); t_(Ret)=30.49 min (race and rac-5)].

[0247] rac-4:

[0248] (1.877 g, 44.3%) (calculated from starting material)

[0249]¹H-NMR (CDCl₃, 360 MHz): δ 1.25-1.86 (10H, m, cyclohexylidene),2.70 (1H, d, J=1.98 Hz, OH), 3.58 (1H, dd, J=7.83, 7.83, 1.98 Hz, H-5),3.69-3,94 (3H, m, H-3, H-4, H-6), 4.17 (1H, dd, J=6.67, 6.56 Hz, H-2),4.38 (1H, dd, J=5.62, 3.64 Hz, H-1), 4.71-5-08 (6H, m, CH ₂Ph),7.22-7.50 (1SH, m, CH₂h. MS: m/z (pos. mode, DCI) 548 [(M+NH₄ ³⁰)⁺,100]. MS: m/z (neg. mode, DCI) 529 [(M−H⁺)⁻, 30]. 439 [(M−C₇H₇ ⁺)⁻, 30].

[0250] rac-5:

[0251] (1.455 g, 34.3%/o) (calculated from starting material)

[0252] Mp: 94.6-95° C. ¹H-NMR (CDCl₃, 360 MHz): δ 1.20-1.88 (10H, m,cyclohexylidene), 2.65 (1H, s, OH), 3.35 (1H, dd, J=9.15, 9.15 Hz, H-5),3.61 (1H, dd, J=9.76, 3.80 Hz, H-3), 3.77 (1H, dd, J=9.21, 6.67 Hz,H-6), 4.12 (1H, dd, J=9.43, 9.43 Hz, H-4), 4.18 (1H, dd, J=6.56, 5.24Hz, H-1), 4.39 (1H, dd, J=4.91, 4.24 Hz, H-2), 4.72-4.99 (6H, m, CH₂Ph), 7.20-7.51 (15H, m, CH₂ Ph). MS: m/z (pos. mode, DCI) 548 [(M+NH₄⁺)⁺, 72]. MS: m/z (neg. mode, DCI 529 [(M−H⁺)⁻, 42]. 439 [(M−C₇H₇ ⁺)⁻,38]. Anal.(C₃₃H₃₈O₆): C calcd, 74.69; found, 74.79. H: cald, 7.22;found, 7.31.

[0253] rac-1,5-Di-O-benzyl-2,3-O-cyclohexylidene-myo-inositol:

[0254] (468 mg, 13.3%) (calculated from starting material)

[0255] Mp: 148.1-148.9° C. ¹H-NMR (CDCl₃, 360 MHz): δ 1.23-1.58 (10H, m,cyclohexylidene), 2.68 (2H, s, OH), 3.38 (1H, dd, J=Hz, H-4), 3.53 (1H,dd, J Hz, H-3), 3.55 (1H, dd, J=Hz, H-6), 3.96 (1H, dd, J=Hz, H-5), 4.09(1H, dd, J Hz, H-1), 4.33 (1H, dd, J=Hz, H-2), 4.63-5.03 (4H, m, CH₂Ph), 7.28-7.47 (10H, m, CH₂ Ph). MS: m/z (pos. mode, DCI) 458 [(M+NH₄⁺)⁺, 100]. MS: m/z (neg. mode, DCI) 439 [(M−H⁺)⁻, 35]. 349 [(M−C₇H₇ ⁺)⁻,73]. HRMS: in/z 439.2120 (M−H⁺)⁻ (calcd for C₂H₃₁O₆, 439.2121).

[0256] rac-1,4,6-Tri-O-benzyl-myo-inositol (rac-6)

[0257] A solution of rac-4 (1.70 g, 3.208 mmol) in MeCN/H₂O (100:1, 3mL) was stirred with trifluoroacetic acid (1 mL) at room temperature for1 h. The solvent was evaporated under reduced pressure. The residue waswashed with cold acetonitrile and the procedure was repeated twice togive the title compound rac-6 (1,095 g, 76%) as a solid.

[0258] Mp: 116,8-117,4° C. ¹H-NMR (CDCl₃, 360 MHz): δ 2.19 (3H, s, OH),3.40-3.52 (2H, m, H-1, H-3), 3.52 (1H, dd, J=9.16, 9.16 Hz, H-5), 3.73(1H, dd, J=9.32, 9.32 Hz, H-6), 3.82 (1H, dd, J=9.37, 9.37 Hz, H-4),4.22 (1H, dd, J=2.82, 2.82 Hz, H-2), 4.68-5.03 (6H, m, CH ₂Ph),7.27-7.43 (15H, m, CH₂ Ph). MS: m/z (pos. mode, DCI) 468 [(M+NH₄ ⁺)⁺,34], 198[(M−3C₇H₇ ⁺+NH₄ ⁺+3H⁺)⁺, 67]. MS: m/z (neg. mode, CI) 449[(M−H⁺)⁻, 28]. 359 [(M−C₇H₇+)⁻, 52]. Anal.(C₂₇H₃₀O₆): C calcd, 71.98;found, 71.66. H: cald, 6.71; found, 6.62.

[0259] Preparation of Enantiomerically Pure1,4,6-Tri-O-benzyl-myo-inositol (6) and 3,4,6-Tri-O-benzyl-myo-inositol(ent-6)

[0260] A solution of the racemic triol (rac-6) (1.069 g, 2.376 mmol) andS-(−)-camphanic acid chloride (1.030 g, 4.752 mmol) in dry pyridine waskept at room temperature for 2 days. All volatile compounds were removedunder reduced pressure and residual pyridine was removed by evaporatingthree times with octane. The residue was dissolved in tert-butyl methylether (100 mL) and was washed twice with sodium hydrogen sulfate (50mL), with phosphate buffer (50 mL), and finally with brine (50 mL). Theorganic layer was dried over Na₂SO₄ and filtered. Evaporation of thesolvent and crystallization from methanol gave pure dia-7 (293 mg, 58%,ee>98%) as a solid. The other diastereomer 7 was isolated and purifiedby preparative HPLC (80% MeOH, 40 mL/min., t_(Ret)=30.49 min) as an oil(214.4 mg, 42%, ee 82%).

[0261] 7: Mp: 187.5-189.8° C. [α]²⁰ _(D)+6,24 (c=1.01 in CHCl₃). ¹H-NMR(CDCl₃, 360 MHz): δ 0.72 (3H, s, CH₃-camph.), 0.91 (3H, s, CH₃-camph.),0.95 (3H, s, CH₃-camph.), 0.99 (3H, s, CH₃-camph.), 1.03 (3H, s,CH₃-camph.), 1.10 (3H, s, CH₃-camph.), 1.49-1.95 (6H, m, CH₂-camph.),2.13-2.44 (3H, m, CH₂-camph.), 3.66 (1H, dd, J=9.49, 2.58 Hz, H-1), 4.07(1H, dd, J=9.68, 9.68 Hz, H-6), 4.28 (1H, dd, J=10.07, 10.07 Hz, H-4),4.33 (1H, dd, J=2.38, 2.38 Hz, H-2), 4.61-4.97 (6H, m, CH ₂Ph), 5.02(1H, dd, J=10.22, 2.63 Hz, H-3), 5.36 (1H, dd, J=9.78, 9.78 Hz, H-5),7.18-7.41 (15H, m, CH₂ Ph. MS: m/z (pos. mode, DCI) 828 [(M+NH₄ ⁺)⁺,70].

[0262] MS: m/z (neg. mode, DCI) 719 [(M−C₇H₇ ⁺)⁻, 8]. Anal.(C₄₉H₆₂O₁₀):C calcd, 72.57; found, 68.86. H: cald, 7.71; found, 7.60.

[0263] dia-7: Mp: 185.3-186.8° C. [α]²⁰ _(D)+2,62 (c=1.03 in CHCl₃).¹H-NMR (CDCl₃, 360 MHz): δ 0.72 (3H, s, CH₃-camph.), 0.85 (3H, s,CH₃-camph.), 0.91 (3H, s, CH₃-camph.), 1.02 (3H, s, CH₃-camph.), 1.07(3H, s, CH₃-camph.), 1.09 (3H, s, CH₃-camph.), 1.46-2.04 (6H, m,CH₂-camph.), 2.06-2.39 (3H, m, CH₂-camph.), 3.68 (1H, dd, J=9.39, 2.61Hz, H-3), 4.04 (1H, dd, J=9.63, 9.63 Hz, H-4), 4.26 (1H, dd, J=10.02,10.02 Hz, H-6), 4.39 (1H, dd, J=2.71, 2.71 Hz, H-2), 4.60-4.94 (6H, m,CH ₂Ph), 4.98 (1H, dd, J=10.31, 2.66 Hz, H-1), 5.35 (1H, dd, J=9.83,9.83 Hz, H-5), 7.15-7.37 (15H, m, CH₂ Ph. MS: m/z (pos. mode, DCI) 828[(M+NH₄ ⁺)⁺, 100].

[0264] MS: m/z (neg. mode, CI) 719 [(M−C₇H₇ ⁺)⁻, 40]. Anal.(C₄₉H₆₂O₁₀):C calcd, 72.57; found, 69.72. H: cald, 7.71; found, 6.89.

[0265] To generate the enantiomerically pure compounds 6 and ent-6 eachester was dissolved in MeOH, 2N KOH (to pH z 14) was added, and thesolution was stirred overnight at room temperature. After the reactionmixture was neutralized with 1N HCl, the solvent was evaporated underreduced pressure and the residual product was extracted with tert-butylmethyl ether (100 mL). The organic layer was washed with sodium hydrogencarbonate (50 mL), phosphate buffer (50 mL) and finally twice with brine(50 mL), dried with Na₂SO₄ and filtered. Evaporation of the solvent gavethe title compounds 6 (99.4 mg, 99%) and ent-6 (142.8 mg, 95%).

[0266] 6: Mp: 117.5-118.2° C. [α]²⁰ _(D)+0.5 (c=1.07 in CHCl₃). ¹H-NMR(CDCl₃, 360 MHz): δ 2.47-2.62 (3H, s, OH), 3.42-3.59 (3H, m, H-1, H-3,H-5), 3.73 (1H, dd, J=9.28, 9.28. Hz, H-6), 3.82 (1H, dd, J=9.23, 9.23Hz, H-4), 4.23 (1H, dd, J=2.64, 2.64 Hz, H-2), 4.67-5.05 (6H, m, CH₂Ph), 7.23-7.53 (15H, m, CH₂ Ph). MS: m/z (pos. mode, DCI) 468 [(M+NH₄⁺)⁺, 100]. MS: m/z (neg. mode, DCI) 449 [(M−H⁺)⁻, 10]. 359 [(M−C₇H₇ ⁺)⁻,56]. HRMS: m/z 449.1960 (M−H⁺)⁻ (calcd for C₂₇H₂₉O₆, 449.1964).

[0267] ent-6: Mp: 116.8-118.9° C. [α]²⁰ _(D)−0.7 (c 1.11 in CHCl₃). ¹Hand MS data were in accordance with those obtained for 7.

[0268] 1,4,6-Tri-O-benzyl-2,3,5-tri-O-butryl-myo-inositol (8).

[0269] A solution of 6 (97.9 mg, 218 μmol), butyric anhydride (213 μL,1.305 mmol), and DMAP (3 mg, 22 μmol) in dry pyridine (4 mL) was stirredat room temperature for 12 h. The solvent was evaporated under highvacuum to give an oil. Residual pyridine was removed by evaporatingthree times with octane. The residue was dissolved in tert-butyl methylether (100 mL) and was washed twice with phosphate buffer (50 mL), withsodium hydrogen sulfate (50 mL), again with phosphate buffer (50 mL),and finally with brine (50 mL). The organic layer was dried over Na₂SO₄and filtered. The solvent was removed under reduced pressure and theresidual oil was purified by preparative HPLC (91% MeOH; 40 mL/min.;t_(Ret)=23.75 min) to give the fully protected compound 8 (105.9 mg,74%).

[0270] [α]²⁰ _(D)+0.50 (c=1.26 in CHCl₃). ¹H-NMR (CDCl₃, 360 MHz): δ0.88 (3H, t, J=7.44 Hz, CH₃-Bt), 0.91 (3H, t, J=7.35 Hz, CH₃-Bt), 1.01(3H, t, J=7.40 Hz, CH₃-Bt), 1.46-1.78 (6H, m, β-CH₂-Bt), 2.05-2.25 (4H,m, α-CH₂-Bt), 2.41 (2H, t, J=7.25 Hz, α-CH₂-Bt), 3.69 (1H, dd, J=9.63,2.63 Hz, H-1), 3.80 (1H, dd, J=9.39, 9.39 Hz, H-6), 3.90 (1H, dd,J=9.97, 9.97 Hz, H-4), 4.43-4.90 (6H, m, CH ₂Ph), 4.96 (1H, dd, J=10.32,2.82 Hz, H-3), 5.22 (1H, dd, J=9.54, 9.54 Hz, H-5), 5.80 (1H, dd,J=2.77, 2.77 Hz, H-2), 7.17-7.38 (15H, m, CH₂ Ph). MS: m/z (pos. mode,DCI) 678 [(M+NH₄ ⁺⁾ ⁺, 100]. MS: m/z (neg. mode, DCI) 659 [(M−H⁺)⁻, 30].569 [(M−C₇H₇ ⁺)⁻, 100]. HRMS: m/z 659.3220 (M−H⁺)⁻ (calcd for C₃₉H₄₇O₉,659.3220).

[0271] 3,4,6-Tri-O-benzyl-1,2,5-tri-O-butyl-myo-inositol (ent-8).

[0272] Compound ent-6 (122.6 mg, 272 μmol) was butyrylated as describedfor 9 to give the fully protected inositol ent-8 (140.8 mg, 78%) as anoil. [α]²⁰ _(D)−0.30 (c=1.35 in CHCl₃). ¹H and MS data were inaccordance with those obtained for 8.

[0273] 2,3,5-Tri-O-butryl-myo-inositol (9).

[0274] Compound 8 (100.7 mg, 153 μmol) was suspended in acetic acid (4mL) and hydrogenated with palladium (10%) on charcoal (92 mg, 915 μmol)under a hydrogen atmosphere in a self-built hydrogenation apparatus for12 h. The catalyst was, removed by ultrafiltration and the filtrate wasfreeze-dried to give 9 (56 mg, 94%) as a hygroscopic solid.

[0275] [α]²⁰ _(D)+15.1 (c 0.67 in CHCl₃). ¹H-NMR (CDCl₃, 360 MHz): δ0.88-1.04 (9H, m, CH₃-Bt), 1.52-1.80 (6H, m, β-CH₂-Bt), 2.22-2.48 (6H,m, α-CH₂-Bt), 2.49-2.74 (3H, s, OH), 3.79-3.86 (2H, m, H-1, H-6), 3.92(1H, dd, J=9.98, 9.98 Hz, H-4), 4.89-5.01 (2H, m, H-3, H-5), 5.56 (1H,dd, J=1.92, 1.92 Hz, H-2). MS: m/z (pos. mode, DCI) 408 [(M+NH₄ ⁺)⁺,100]. MS: m/z (neg. mode, DCI) 196 [(M−3Bt⁺+H₂O+H⁺)²⁻, 100].

[0276] HRMS: m/z 389.1800 (M−H⁺)⁻ (calcd for C₁₈H₂₉O₉, 389.1812).

[0277] 1,2,5-Tri-O-butyryl-myo-inositol (ent-9).

[0278] Compound ent-8 (134.5 mg, 204 μmol) was hydrogenated as describedfor 8 to give ent-9 (74.7 mg, 94%) as a hygroscopic solid.

[0279] [α]²⁰ _(D)−14.5 (c=0.42 in CHCl₃). ¹H and MS data were inaccordance with those obtained for 9.

[0280] 2,3,5-Tri-O-butyryl-myo-inositol 1,4,6-Tris(dibenzyl)phosphate(10).

[0281] A solution of tetrol 9 (40.6 mg, 104 μmol) and tetrazole (55 mg,781 mmol) in acetonitrile (5 mL) was treated with dibenzylN,N-diisopropylphosphoramidite (257 μL, 781 μmol) for 20 h andsubsequently oxidized with peracetic acid (200 μL, 781 μmol) at −40° C.After the mixture warmed to room temperature, the solvent was removedunder reduced pressure and the residual oil was purified by preparativeHPLC (88% MeOH; 40 mL/min., t_(Ret)=34.75 min) to give the fullyprotected compound 10 (96 mg, 79%) as a clear oil.

[0282] [α]²⁰ _(D)−5.5 (c=8 in CHCl₃). ¹H-NMR (CDCl₃, 360 MHz): δ 0.69(3H, t, J=7.48 Hz, CH₃-Bt), 0.81 (3H, t, J=7.38 Hz, CH₃-Bt), 0.96 (3H,t, J=7.38 Hz, CH₃-Bt), 1.24-1.75 (6H, m, β-CH₂-Bt), 1.98-2.40 (6H, m,α-CH₂-Bt), 4.47 (1H, ddd, J=9.78, 8.46, 2.69 Hz, H-1), 4.73-5.13 (15H,m, H-3, H-4, H-6, 6×CH ₂Ph), 5.32 (1H, dd, J=9.59, 9.59 Hz, H-5), 5.87(1H, dd, J=2.59, 2.59 Hz, H-2), 7.16-7.38 (30H, m, 6×CH₂ PH). ³¹P-NMR(CDCl₃, ¹H decoupled, 145.8 MHz): −0.69 (1 P, s), −0.55 (1 P, s), −0.42(1 P, s). MS: m/z (neg. mode, DCI) 1079 [(M−C₇H₇ ⁺)⁻, 100]. HRMS: m/z1079.3449 (M−C₇H₇ ⁺)⁻(calcd for C₅₃H₆₂O₁₈P₃, 1079.3149).

[0283] 1,2,5-Tri-O-butyryl-myo-inositol 3,4,6-Tris(dibenzyl)phosphate(ent-10).

[0284] Tetrol ent-9 (63.6 mg, 163 μmol) was phosphorylated as describedfor compound 10 to give the fully protected phosphate ent-10 (157.2 mg,82%) as a clear oil. [α]²⁰ _(D)+5.43 (c=10 in CHCl₃).

[0285]¹H, ³¹P and MS data were in accordance with those obtained for 10.

[0286] 2,3,5-Tri-O-butyryl-myo-inositol 1,4,6-Trisphosphate (11).

[0287] Compound 10 (94.1 mg, 80 μmol) was hydrogenated with palladium(10%) on charcoal as described for compound 9 to give the title compound11 (58.8 mg, 100%) as a hygroscopic solid after freeze-drying.

[0288] [α]²⁰ _(D)−1.71 (c=1.49 in CHCl₃). ¹H-NMR (D₂O, 360 MHz): δ 0.74(3H, t, J=7.61 Hz, CH₃-Bt), 0.78 (3H, t, J=7.42 Hz, CH₃-Bt), 0.83 (3H,t, J=7.53 Hz, CH₃-Bt), 1.32-1.65 (6H, m, β-CH₂-Bt), 2.15-2.44 (6H, m,α-CH₂-Bt), 4.32-4.59 (3H, m, H-1, H-4, H-6), 5.07 (1H, dd, J=9.92, 2.65Hz, H-3), 5.14 (1H, dd, J=9.52, 9.52 Hz, H-5), 5.57 (1H, dd, J=2.46,2.46 Hz, H-2). ³¹P-NMR (D₂O, ¹H decoupled, 145.8 MHz): δ 0.09 (1P, s),0.70 (1P, s), 0.78 (1P, s).

[0289] MS: m/z (neg. mode, DCI) 629 [(M−H⁺)⁻; 28], 559 [(M−Bt⁺)⁻, 52].

[0290] 1,2,5-Tri-O-butyryl-myo-inositol 3,4,6-Trisphosphate (ent-11).

[0291] Compound ent-10 (155.3 mg, 133 μmol) was hydrogenated asdescribed for 11 to give the free acid ent-11 (90.7 mg, 100%) as ahygroscopic solid. [α]²⁰ _(D)+1.85 (c 2.75 in CHCl₃). ¹H, ³¹P and MSdata were in accordance with those obtained for

[0292] 2,3,5-Tri-O-butyryl-myo-inositol 1,4,6-TrisphosphateHexakis(acetoxymethyl) Ester

[0293] DIEA (118 μL, 690 μmol) and acetoxymethyl bromide (69 μL, 690mmol) were added to a suspension of compound 11 (28.7 mg, 46 μmol) indry acetonitrile (2 mL) under an argon atmosphere. After the reactionmixture stirred in the dark for 5 days, all volatile compounds wereremoved under reduced pressure and the crude residue was purified bypreparative HPLC (70% MeOH; 40 mL/min.; t_(Ret)=18.20 min) to givecompound 1 (34.3 mg, 71%) as a clear syrup. [α]²⁰ _(D)+0.44 (c 1.127 intoluene).

[0294]¹H-NMR (toluene-d₈, 360 MHz): δ 0.85 (3H, t, J=7.68 Hz, CH₃-Bt),0.89 (3H, t, J=7.43 Hz, CH₃-Bt), 1.06 (3H, t, J=7.48 Hz, CH₃-Bt),1.43-1.96 (20H, m, β-CH₂-Bt, 6×OAc), 2.00-2.49 (8H, m, α/β-CH₂-Bt),2.64-2.76 (2H, m, α-CH₂-Bt), 4.56 (1H, ddd, J=9.54, 9.54, 2.84 Hz, H-1),4.94 (1H, dd, J=9.63, 9.63 Hz, H-6), 5.02 (1H, dd, J=10.17, 2.84 Hz,H-3), 5.03 (1H, dd, J=9.73, 9.73 Hz, H-4), 5.29-5.80 (13H, m, H-5,6×CH₂OAc), 5.96 (1H, dd, J=2.64, 2.64 Hz, H-2). ³¹P-NMR (toluene-d₈, ¹Hdecoupled, 145.8 MHz): 6-3.72 (1P, s), −3.40 (1P, s), −3.34 (1P, s). MS:m/z (neg. mode, DCI) 989 [(M−CH₂OCOCH₃ ⁺)⁻, 100]. HRMS: m/z 989.1858(M−CH₂OCOCH₃ ⁺)⁻ (calcd for C₃₃H₅₂O₂₈P₃, 989.1858).

[0295] 1,2,5-Tri-O-butyryl-myo-inositol 3,4,6-TrisphosphateHexakis(acetoxymethyl) Ester (ent-1).

[0296] The free acid ent-11 (49.4 mg, 78 μmol) was alkylated asdescribed for compound 1 to give the acetoxymethyl ester ent-1 (36.8 mg,44%) as a clear syrup. [α]²⁰ _(D)−0.41 (c=1.227 in toluene). ¹H, ³¹p andMS data were in accordance with those obtained for 1.

[0297] myo-Inositol 1,4,6-Trisphosphate (2)

[0298] Compound 11 (22,4 mg, 36 μmol) was treated with 0.5 N KOH (2 mL)to adjust the pH value to. 14. The solution was stirred at roomtemperature for 4 days. The reaction mixture was directly poured onto anion-exchange column (Dowex 50 WX 8, H⁺) for purification. Lyophilizationgave compound 2 (5,8 mg, 39%).

[0299] [α]20_(D)−8.49 (c=0.48 in MeOH). ¹H-NMR (D₂O+6 eq. DIEA, 360MHz): δ 3.35 (1H, dd, J=9.05, 9.05 Hz, H-5), 3.41-3.64 (1H, m, H-3,DIEA), 3.73 (1H, ddd, J=9.87, 9.87, 2.52 Hz, H-1), 3.90-4.11 (2H, m,H-4, H-6), 4.14 (1H, dd, J=2.33, 2.33 Hz, H-2). ³¹P-NMR (D₂O+6 eq. DIEA,¹H decoupled, 145.8 MHz): δ 4.32 (1P, s), 5.41 (1P, s), 5.67 (1P, s).MS: m/z (neg. mode, DCI) 159 [(M−3[PO(OH)₂]⁻-H₂O)³⁻; 90].

[0300] myo-Inositol 3,4,6-Trisphosphate (ent-2)

[0301] The free acid ent-11 (41,2 mg, 65 μmol) was saponified asdescribed for compound 2 (12,7 mg, 46%). [ ]²⁰ _(D)+4.49 (c=0.85 inMeOH). ¹H, ³¹P, and MS data were in accordance with those obtained for2.

EXAMPLE 2 Synthesis of rac-(3aS, 4R, 5S, 6R, 7S,7aS)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol(24)

[0302]

[0303] Referring to the reaction scheme shown in FIG. 4, the compoundrac-(3aS, 4R, 5S, 6R, 7S,7aS)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol(24) was synthesized as follows:

[0304] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-6-allyloxy-2,3,4,5-tetrabenzyloxy-1-hydroxy-1-propyl-cyclohexane(20):

[0305] 0.305 g Magnesium powder (12.6 mmol) was suspended in 15 mL drydiethyl ether under an argon atmosphere and 1.25 mL iodopropane (12.9mmol) was added. After the initial boiling stopped the mixture washeated to reflux for another 15 min. In a second reaction vessel 0.83 ginosose 2 (1.43 mmol) was suspended in 10 mL dry diethyl ether and theGrignard mixture was added in portions under vigorous stirring andargon. After there was no starting material detectable by analyticalHPLC (RP18, 1.5 mL/min, 90% MeOH, t_(Ret)=6.35 min) the reaction wasquenched by adding water. The organic phase was diluted witht-butylmethyl ether and was washed with Na₂SO₄-solution (10%), 0.5 Mphosphate buffer pH=7, and brine. The organic phase was dried overNa₂SO₄, filtered, and dried under reduced pressure. The residue waspurified by preparative HPLC (RP18, 89% MeOH, 40 mL/min). 0.374 g(t_(Ret)=37.10 min, 0.60 mmol, 42% yield) of the equatorial alkylatedproduct 20 was isolated as a colorless solid as well as 0.113 g(t_(Ret)=45.45 min, 0.20 mmol, 14.6% yield) of the axial alkylatedepimner.

[0306] Equatorially alkylated epimer 20:

[0307] m.p.: 103.5° C.

[0308]¹H-NMR (200 MHz, CDCl₃): δ 7.51-7.28 (m, 20H, Ph), 6.18-5.96 (m,1H, OCH₂CHCH₂), 5.43-5.20 (m, 2H, OCH₂CHCH₂), 5.194.67 (m, 8H, CH₂Ph),4.63-4.51 and 4.30-4.17 (m, 2H. OCH₂C₂H₃), 4.17 (dd, 1H, ³J=9.7 Hz,H-5),4.12 (dd, 1H, ³J=9.7 Hz, H-3), 3.61 (dd, 1H, ³J=9.6 Hz, H-4), 3.51(d, 1H, ³J=9.3 Hz, H-6), 3.39 (d, 1H, ³J=9.3 Hz, H-2), 2.29 (br s, 1H,OH), 2.00-1.74 (m, 2H, CH₂CH₂CH₃), 1.50-1.08 (m, 2H, CH₂CH₂CH₃), 0.97(t, 3H, CH₂CH₂CH₃, ³J=7.1 Hz, CH₃)

[0309] MS (neg. mode, FAB, NBA) m/z (%): 621 [M−H⁺] (38), 531 [M−Bn⁺](100)

[0310] MS (pos. mode, FAB, NBA) m/z (%): 640 [M+NH₄ ⁺] (100)

[0311] C₄₀H₄₆O₆ (622.80): calcd. C 77.14, H 7.44; found C 77.20, H 7.30

[0312] Axially alkylated epimer:

[0313]¹H-NMR (200 MHz, CDCl₃): δ 7.56-7.21 (m, 20H, Ph), 6.17-5.94 (m,1H, OCH₂CHCH₂), 5.46-5.20 (m, 2H, OCH₂CHCH₂), 5.04-4.86 (m, 4H, CH₂Ph),4.55-4.30 (OCH₂CHCH₂), 3.79-3.32 (m, 5H, H-2, H-3, H-4, H-5), 2.52 (brs, 1H, OH), 1.90-1.63 (m, 4H, CH₂CH₂), 1.13-0.97 (m, 3H, CH₃)

[0314] MS (neg. mode, FAB, NBA) m/z (%): 621 [M−H⁺] (4), 531 [M−Bn⁺+H³⁰](2)

[0315] MS (pos. mode, FAB, NBA) m/z (%): 623 [M+H⁺] (<1), 531 [Bn⁺](100)

[0316] Synthesis of rac-(1S, 2R, 3S, 4R, 5R,6S)-2,3,4,5-tetrabenzyloxy-1,6-dihydroxy-1-propyl-cyclohexane (21)

[0317] 20 (0.30 g, 0.48 mmol) was dissolved in 20 mL aqueous ethanol(90%). 93 mg (0.101 mmol) Wilkinson catalyst and 50 μL diisopropylethylamine were added and the mixture was heated to reflux for 12 hours.After cooling to room temperature 30 μL triflouro acetic acid was addedand the solution was allowed to stir for 12 days by when the startingmaterial had disappeared. The solution was dried in high vacuum and theresidue was dissolved in t-butylmethyl ether and washed with 0.5 Mphosphate buffer pH=7 and brine. The organic phase was filtered afterbeing dried over Na₂SO₄ and the filtrate was dried under reducedpressure. Further purification was accomplished by preparative HPLC (89%MeOH, 40 m/min, t_(Ret)=21.40 min) to give 220 mg of 21 (0.38 mmol,78.4% yield) as a colorless oil.

[0318]¹H-NMR-(200 MHz, d₆-DMSO): δ 7.43-7.23 (m, 20H, Ph), 5.034.58 (m,8H, CH₂-Bn), 4.02 (dd; 1H, ³J=9.4 Hz, H-5), 3.82 (dd, 1H, ³J=9.3 Hz,H-3), 3.52 (dd, 1H, ³J=9.6 Hz, H-4), 3.43 (d, 1H, ³J=9.4 Hz, H-2), 3.43(dd, 1H, ³J=9.4 Hz, 5.1 Hz, H-6), 2.27 (d, 1H, ³J=5.1 Hz, OH), 2.18 (brs, 1H, OH), 1.87-1.59 (m, 2H, CH(Pr)), 1.47-0.94 (m, 2H, CH(Pr)), 0.87(t, 3H, ³J=7.1 Hz, CH₃)

[0319] MS (neg. mode, FAB, NBA) m/z (%): 581 [M−H⁺] (17), 491 [M−Bn⁺](4)

[0320] Synthesis of rac-(3aR, 4R, 5S, 6S, 7R,7aS)-4,5,6,7-tetrabenzyloxy-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol(22)

[0321] 21 (0.22 g, 0.37 mmol) was dissolved in 20 mL dry acetone. 5 mL(40.8 mmol) 2.2-dimethoxypropane and 48 mg (0.25 mmol) p-toluenesulfonic acid were added an the solution was stirred for three days atroom temperature. Then the reaction was stopped by adding 150 μL (1.08mmole) triethylamine. All volatile components were removed under reducedpressure and the residue was dissolved in t-butylmethyl ether and washedwith 0.5 M phosphate buffer pH=7 and brine. The organic phase was driedover Na₂SO₄, filtered and dried under reduced pressure. The residue waspurified by preparative HPLC (RP18, 94% MeOH, 40 mL/min, t_(Ret)=35.10min) to give 22 (160 mg, 0.25 mmol, 69.3% yield) as a slightly yellowoil.

[0322]¹H-NMR (200 MHz, CDCl₃): δ 7.52-7.18 (m, 20H, Ph), 4.85-4.49 (m,8H, CH₂-Ph), 4.09 (dd, 1H, ³J=5.3 Hz, H-4), 4.01 (dd, ³J=7.3 Hz, ³J=0.3Hz, H-7), 3.94 (dd, ³J=6.9 Hz, ³J=5.3 Hz, H-5), 3.67 (dd, ³J=7.0 Hz,H-6), 3.63 (dd, ³J=5.3 Hz, H-7a), 1.73-1.61 (m, 2H, CH(Pr)), 1.38-1.17(m, 2H, CH(Pr)), −1.51 (s, 3H, CH₃), 1.42 (s, 3H, CH₃), 0.79 (t, 3H,³J=7.3 Hz, CH₃Pr))

[0323] MS (pos. mode, DCI, NH₃) m/z (%): 640 [M+NH₄ ⁺] (17)

[0324] MS (neg. mode, DCI, NH₃) m/z (%): 621 [M−H⁺] (2), 531 [M−Bn⁺](28)

[0325] Synthesis of rac-(3aS, 4R, 5S, 6R, 7S,7aS)-4,5,6,7-tetrakis[[bis(benzyloxy)-phosphoryl]oxy]-2,2-dimethyl-3a-propyl-cis-hexahydrobenzo-(1,3)-dioxol(23)

[0326] 22 (160.8 mg, 0.258 mmol) was dissolved in 10 mL ice-cold ethanol(−20° C.) and palladium on charcoal (10%, 990 mg, 0.93 mmol) was addedas a suspension in 10 mL ice-cold ethanol under argon. The mixture wassubjected to a hydrogen atmosphere in a self-built hydrogenationapparatus. After a week of vigorous stirring the catalyst was filteredoff and the filtrate dried under reduced pressure. The residue and 0.214g tetrazole (3.06 mmol) were dissolved in 7 mL dry acetonitrile. Underargon 0.814 mL (2.42 mmol) dibenzyl N,N-diisopropyl phosphoramidite wasadded and the mixture was stirred for 48 hours before it was cooled −40°C. and 0.5 mL peracetic acid was carefully added. The solution wasallowed to warm to room temperature and the volatile components wereremoved in high vacuum. Further purification by preparative HPLC (93%MeOH, 40 mL/min, t_(Ret)=21.5 min) gave 23 (36.7 mg, 22 μmol, 8.6%yield) as a colorless oil.

[0327]¹H-NMR (200 MHz, CDCl₃): δ 7.39-7.12 (m, 40H, Ph), 0.72 (t, 3H,CH₃), 5.27-4.76 (m, 20H, CH₂-Ph, H-4, H-5, H-6, H-7), 4.24 (d, 1H,³J=5.2 Hz, H-7a), 1.89-1.66 (m, 2H, CH(Pr)), 1.48 (s, 3H, CH₃),1.48-1.22 (m, 2H, CH(Pr)), 1.32 (s, 3H, CH₃), 0.73 (t, 3H, ³J=7.2 Hz,CH₃(Pr))

[0328]³¹P-NMR (81 MHz, CDCl₃): δ −0.28 (2P), −0.78 (P), −0.98 (P)

[0329] MS (pos. mode, DCI, NH₃) m/z (%): 1303 [M+H⁺] (100)

[0330] MS (neg. mode, DCI, NH₃) m/z (%):1301 [M−H⁺] (9), 1210 [M−H⁺−Bn](100), 1121 [M−2*Bn⁺+H⁺] (49)

[0331] Synthesis of rac-(3aS, 4R, 5S, 6R, 7S,7aS)4,5,6,7-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol(24)

[0332] Under an argon atmosphere 500 mg palladium on charcoal (10%, 0.47mmol) were carefully suspended in 5 mL ice-cold ethanol (−20° C.) and0.1 mL diisopropylethyl amine was added. 26 mg (20 μmol) of phosphorustriester 23 was added dissolved in 10 mL ice-cold ethanol. The mixturewas subjected to a hydrogen atmosphere in a self-built hydrogenationapparatus. After a week of vigorous stirring the catalyst was filteredoff and the filtrate dried under reduced pressure. The residue wasdissolved in 1 mL dry acetonitrile under an argon atmosphere and 132 μldiisopropylethyl amine (98 μmole) and 178 μl acetoxymethyl bromide (178μmol) were added. After 48 hours all volatile components were removed inhigh vacuum and the residual oil was subjected to preparative HPLC togive 9.2 mg of 24 (8 μmol, 39.7% yield) as a colorless oil.

[0333]¹H-NMR (200 MHz, d₈-toluene): δ 5.93-5.51 (m, 16H, OCH₂O),5.47-5.28 (m, 2H, H-5, H-7), 5.17-5.97 (m, 2H, H-4, H-6), 4.24 (d, 1H,³J=4.8 Hz, H-7a), 1.90 (s, 3H, OAc), 1.81 (s, 3H, OAc), 1.81 (2s, 6H,OAc), 1.78 (s, 3H, OAc), 1.76 (s, 3H, OAc), 1.76 (s, 3H, OAc), 1.75 (s,3H, OAc), 1.70-1.51 (m, 2H, CH(Pr)), 1.54 (s, 3H, CH₃), 1.41-1.28 (m,2H, CH(Pr)), 1.18 (s, 3H, CH₃), 1.03 (t, 3H, ³J=6.9 Hz, CH₃ (Pr))

[0334]³¹P-NMR (81 MHz, d₈-toluene): δ −2.55 (2P), −3.34 (2P)

[0335] MS (pos. mode, FAB, NBA) m/z (%):1181 [M+Na⁺] (5)

[0336] MS (neg. mode, FAB, NBA) m/z (%): 1085 [M−CH₂OAc⁺] (6)

[0337] HR-MS: calcd. for C₃₃H₅₃O₃₂P₄ [M−CH₂OAc⁺] 1085.1471, found1085.1468

EXAMPLE 3 Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane(29)

[0338]

[0339] Referring to the reaction scheme shown in FIG. 5, the compoundrac-(1S, 2R, 3S, 4R, 5S,6S)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane(29) was synthesized as follows:

[0340] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-6-allyloxy-2,3,4,5-tetrabenzyloxy-1-butyroxy-1-propyl-cyclohexane(25)

[0341] 348 mg (0.56 mmol) 20 were dissolved in 25 mL dry toluene. Underan argon atmosphere 2.5 mL (24 mmol) butyric acid anhydride and 250 μLphosphazene base-P₁-t-Bu (0.98 mmol) were added and the solution wasrefluxed for 18 hours. After cooling the solution was diluted witht-butylmethyl ether. The organic layer was extracted twice withphosphate buffer (0.5 M, pH 7), followed by brine, then dried overNa₂SO₄ and filtered. The solvent was removed under reduced pressure andthe solid residue was recrystallized from methanol to give 25 (316 mg,0.46 mmol, 81.4% yield) as a colorless solid.

[0342] m.p.: 115° C.

[0343]¹H-NMR (200 MHz, CDCl₃): δ 7.41-7.19 (m, 20H, Ph), 6.07-5.83 (m,1H, CH₂CHCH₂), 5.31 (ddt, 1H, ³J=17.2 Hz, ²J=³J=⁴J=1.6 Hz, CH₂CHCHH),5.16 (ddt, 1H, ³J=10.5 Hz), 5.03-4.69 (m, 8H, CH₂Ph), 4.52-4.10 (m, 2H,CH₂ CHCH₂), 3.95 (dd, 1H, ³J=9.4 Hz, H-3), 3.91 (dd, 1H, ³J=9.4 Hz,H-5), 3.52 (dd, 1H, ³J=9.5 Hz, H-4), 3.44 (d, 1H, ³J=9.4 Hz, H-6), 3.38(d, ³J=9.4 Hz, H-1), 2.80-2.51 (m, 2H, α-CH₂(Bt)), 2.34 (t, 2H, ³J=7.3Hz, C(OBt)CH₂), 1.82-1-54 (m, 2H, CH₂(Bt)), 1.39-1.10 (m, 2H,C(OBt)CH₂CH₂), 1.01 (t, 3H, ³J=7.2 Hz, CH₃), 0.90 (t, 3H, ³J=7.0 Hz,CH₃)

[0344] MS (negative mode, DCI, NH₃) m/z (%): 691 [M−H⁺] (7), 651[M−C₃H₅+] (2), 621 [M−Bt⁺] (6), 601 [M−Bn⁺] (17), 511 [M−2*Bn⁺+H⁺] (3)

[0345] C₄₄H₅₂O₇ (692.89): calcd. C 76.27H 7.56; found C 76.49H 7.41

[0346] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5-tetrabenzyloxy-1-butyroxy-6-propoxy-1-propyl-cyclohexane(26)

[0347] The allyl ether 21 (182 mg, 0.26 mmol) was dissolved in a mixtureof 7 mL ethanol and 1 mL acetone and 18 mg Wilkinson catalyst (19 μmol)was added. The mixture was stirred in a hydrogen atmosphere for 3 days,before the solvents were removed under reduced pressure. The residue waschromatographed by preparative HPLC (95% MeOH, 40 mL/min, t_(Ret)=35.00min) to give 26 as a colorless solid(108 mg, 0.16 mmol; 59.2% yield).

[0348] m.p.: 114.5° C.

[0349]¹H-NMR (200 MHz, CDCl₃): δ 7.42-7.18 (m, 20H, Ph), 5.03-4.78 (m,8H, CH₂Ph), 3.97-3.85 (m, 1H, OCHHCH₂), 3.94 (dd, 1H. ³J=9.4 Hz, H-5),3.88 (dd, 1H, ³J=9.4 Hz, H-3), 3.54-3.44 (m, 1H, OCHHCH₂), 3.50 (dd, 1H,³J=9.6 Hz, H-4), 3.46 (d, 1H, ³J=9.5 Hz, H-2), 3.33 (d, 1H, ³J=9.4 Hz,H-6), 2.82-2.51 (m, 2H, α-CH₂(Bt)), 2.33 (t, 2H, ³J=7.3 Hz, C(OBt)CH₂),1.80-1.52 (m, 5H, CH₂), 1.38-1.13 (m, 3H, CH₂), 1.01 (t, 3H, ³J=7.3 Hz,CH₃), 0.95 (t, 3H, ³J=7.2 Hz, CH₃), 0.91 (t, 3H, J=7.2 Hz, CH₃)

[0350] MS (pos. mode, DCI, NH₃) m/z (%): 712 [M+NH₄ ⁺] (23), 695 [M+H⁺](7), 91 [Bn⁺] (100),

[0351] MS (neg. mode, DCI, NH₃) m/z (%): 603 M−Bn⁺ (100)

[0352] C₄₄H₅₄O₇ (694.91) calcd. C 76.05, H 7.83; found C 76.18, H 7.98

[0353] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5-tetrakis[[bis(benzyloxy)-phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane(27a)

[0354] 107 mg Palladium on charcoal (10%, 0.10 mmol) was added to asolution of 26 (95 mg, 0.16 mmol) in 7.5 mL glacial acetic acid. Themixture was kept under a hydrogen atmosphere for 12 hours and wasstirred vigorously. Subsequently, the catalyst was removed byultrafiltration and the filtrate was freeze-dried. The residue wasdissolved in 3 mL acetonitrile under an argon atmosphere and 0.5 mLdibenzyl N,N-diisopropyl phosphoramidite (1.49 mmol) und tetrazole (117mg, 1.67 mmol) were added. The mixture was stirred at room temperaturefor 12 hours, then cooled to −40° C. and treated with peracetic acid(32% w/w in acetic acid, 0.350 mL, 1.67 mmol). After the mixture reachedroom temperature all volatile components were removed in high vacuum andthe residue was purified by preparative HPLC (90% MeOH, 40 mL/min). Thechromatogram showed two major products. Accordingly, axiallyphosphorylated compound 27b (84 mg, 42% yield, t_(Ret)=48.3 min) andequatorially phosphorylated compound 27a (68 mg, 34% yield, t_(Ret)52.0min) were isolated as colorless oils.

[0355]¹H-NMR (200 MHz, CDCl₃): δ 7.32-7.12 (m, 40H, Ph), 5.20-4.54 (m,20H, CH₂-Ph, H-2, H-3, H-4, H-5), 3.84-3.68 (m, 1H, OCHHCH₂CH₃) 3.73 (d,³J=7.0 Hz, H-6), 3.36-3.23 (m, 1H, OCHHCH₂CH₃), 2.69-2.25 (m, 2H,α-CH₂(Bt)), 2.20. (t, 2H, C(OBt)CH₂), 1.67-1.31 (m, 6H, CH₂), 0.88 (t,3H, ³J=7.2 Hz, CH₃), 0.80 (t, 3H, ³J=7.5 Hz, CH₃) 0.76 (t, 3H, ³J=7.2Hz, CH₃)

[0356]³¹P-NMR (81 MHz, CDCl₃): δ 0.32 (1P), −0.16 (1P), −0.16 (1P),−0.47 (1P)

[0357] MS (neg. mode, DCI, NH₃) m/z (%): 1373 [M−H⁺] (1), 1284 [M−Bn⁺](16)

[0358] Migration Product 27b:

[0359]¹H-NMR (200 MHz, CDCl₃): δ 7.58-7.10 (m, 40H, Ph), 5.34-4.80 (m,19H, CH₂-Ph, H-2, H-3, H-5), 4.49 (ddd, 1H, J=10.1 Hz, H-4), 4.04-3.89(m, 1H, OCMHCH₂CH₃), 3.44 (dd, 1H, J=9.8 Hz, 3.7 Hz, H-6), 3.32 (m, 1H,OCHHCH₂CH₃), 2.54-2.04 (m, 4H, α-CH₂(Bt), C₁CH₂), 1.90-1.08 (m, 6H),0.94 (t, 3H, ³J=6.9 Hz, CH₃), 0.79 (t, 3H, ³J=7.7 Hz, CH₃), 0.75 (t, 3H,³J=7.0 Hz, CH₃)

[0360]³¹P-NMR (81 MHz, CDCl₃): δ 0.90 (1P), 0.33 (1P), −0.01 (1P), −5.62(1P)

[0361] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-1-butyroxy-4,5,6,7-tetrakis(phosphonooxy)-6-propoxy-1-propyl-cyclohexane(28)

[0362] 27a (73 mg, 53 μmols) was dissolved in 4 mL glacial acetic acidand 130 mg palladium on charcoal (10% Pd, 0.12 mmol) was added. Themixture was kept under a hydrogen atmosphere for 12 hours and wasstirred vigorously. Subsequently, the catalyst was removed byultrafiltration and the filtrate was freeze-dried to give 28 (35 mg, 99%yield) as a colorless oil.

[0363]¹H-NMR (200 MHz, DMSO-d₆): δ 4.53-4.20 (m, 4H, H-2, H-3, H-4,H-5), 3.97-3.81 (m, 1H, OCHH), 3.60 (d, 1H, ³J=9.3 Hz, H-6), 3.41-3.22(m, 1H, OCHH), 2.66-2.23 (m, 2H, α-CH₂(Bt)), 2.17 (t, 2H, ³J=7.3 Hz,C(OBt)CH₂), 1.65-1.35 (m, 4H, CH₂), 1.27-1.05 (m, 2H, CH₂), 0.87 (t, 3H,³J=6.9 Hz, CH₃), 0.85 (t, 3H, ³J=6.5 Hz, CH₃), 0.84 (t, 3H, ³J=7.1 Hz,CH₃)

[0364]³¹P-NMR (81 MHz, DMSO-d₆): δ 0.78 (1P), 0.43 (1P), −0.05 (1P),−0.12 (1P)

[0365] MS (pos. mode, FAB, NBA) m/z (%): 655 [M−H⁺] (<1), 585[M−Bt⁺+2H⁺] (<1)

[0366] MS (neg. mode, FAB, NBA) m/z (%): 653 [M−H⁺] (24), 97 [H₂PO₃ ⁻](27)

[0367] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,65)4,5,6,7-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane(29)

[0368] 23 mg Tetrakisphosphate 28 (35 μmol) was dissolved in 1 mLacetonitrile under an argon atmosphere and 17 μl diisopropylethyl amine(0.69 nmol) and 69 μl acetoxymethyl bromide (0.69 mmol) were added. Thesolution was stirred for 48 hours at room temperature when all volatilecomponents were removed in high vacuum. The residue was purified bypreparative HPLC (40 mL/min, 70% MeOH, t_(Ret)=32.0 min) to give 29 (20mg, 46.7% yield) as a colorless oil.

[0369]¹H-NMR (200 MHz, toluene-d₈): δ 6.02-5.53 (m, 8H, OCH₂O),5.13-4.89 (m, 2H, H-3, H-5), 4.74-4.54 (m, 2H, H-2, H-4), 4.08-3.92 (m,1H, OCHHCH₂CH₃), 3.36-3.19 (m, 1H, OCHHCH₂CH₃), 3.19 (d, 1H, ³J=8.9 Hz,H-6), 3.03-2.54 (m, 2H, α-CH₂(Bt)), 2.35-2.21 (m, 2H, C(OBt)CH₂),1.92-1.53 (m, 6H, CH₂), 1.92 (s, 6H, 2Ac), 1.84 (s, 3H, Ac), 1.81 (s,6H, 2Ac), 1.78 (s, 3H, Ac), 1.75 (s, 3H, Ac), 1.74 (s, 3H, Ac), 1.00 (t,3H, ³J=7.4, CH₃),0.92 (t, 3H, ³J=7.4, CH₃)

[0370]³¹P-NMR (81 MHz, toluene-d₈): δ 2.79 (1P), −3.02 (1P), −3.27 (1P),−3.68 (1P)

[0371] MS (neg. mode, FAB, NBA) m/z (%): 1157 [M−CH₂OAc⁺] (7), 1085 [M−2CH₂OAc⁺+H⁺] (11), 241 [(AcOCH₂O)₂PO₂] (97)

[0372] HR-MS: calcd. for C₃₇H₆₁O₃₃P₄ 1157.2046, found 157.2046

EXAMPLE 4 Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxymethoxy)phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane(34)

[0373] Referring to the reaction scheme shown in FIG. 6, the compoundrac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxymethoxy)phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane(34) was synthesized as follows:

[0374] Synthesis of rac-(1R, 2R, 3S, 4S, 5R,6S)-2,3,4,5-tetra(benzyloxy)-6-butyroxy-1-hydroxy-1-propyl-cyclohexane(31)

[0375] Magnesium powder (0.968 g, 40 mmol) was overlayed with 50 mL drydiethyl ether and 3.8 mL iodopropane (39 mmol) was added. The reactionwas started by adding some iodine crystals and after the boiling slowed,the mixture was held under reflux for 15 min.

[0376] The inosose 30 (0.938 g, 1.54 mmol) was suspended in 20 mLdiethyl ether and 2 mL of the freshly prepared Grignard reagent wasadded under an argon atmosphere. After 3 hours the reaction was quenchedby adding ice and ethyl acetate. The organic layer was washed withaqueous Na₂SO₃ (0.1 M), phosphate buffer (pH 7), and brine. After dryingover Na₂SO₄ and filtration the solvent was removed under reducedpressure. The residue was prepurified by flash chromatography(n-hexane/ethyl acetate 3:1 v/v, silica 60, TLC: R_(f) 0.2). Thecombined product-containing fractions were dried and N was purified bypreparative HPLC (89% MeOH, 40 mL/min, t_(Ret)=35.5 min) to give thecolorless solid 31 (127 mg, 0.20 mmol, 12.6% yield).

[0377] m.p.: 143.3° C.

[0378]¹H-NMR (200 MHz, CDCl₃): 7.57-7.21 (m, 20H, Bn), 5.17 (d, 1H,³J=10.0 Hz, H-6), 5.13-4.69 (m, 8H, CH₂Bn), 4.12 (dd, 1H, ³J=9.4 Hz,H-3), 4.10 (dd, 1H, 3j=9.7 Hz, H-5), 3.70 (dd, 1H, ³J=9.5 Hz, H-4), 3.59(d, 1H, ³J=9.3 Hz, H-2), 2.54-2.19 (m, 2H, α-CH₂(Bt)), 1.92-1.08 (m, 6H,CH₂), 0.99 (t, 3H, ³J=7.3 Hz, CH₃), 0.89 (t, 3H, J=6.9 Hz, CH₃)

[0379] MS (pos. mode, FAB, NBA) in/z (%): 653 [M+H⁺] (1), 635 [M+H⁺−H₂O](3), 91 [Bn⁺]3 (100)

[0380] MS (neg. mode, FAB, NBA) m/z (%): 651 [M−H⁺] (<1), 582 [M−Bt⁺](4)

[0381] C₄₁H₄₈O₇ (652.83): calcd. C 75.43, H 7.41; found C 74.87H 7.30

[0382] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-1,2,3,4,5-6-butyrloxy-pentakis-[[bis(benzyloxy)phosphoryl]oxy]-2-propyl-cyclohexane(32)

[0383] The tetrabenzyl ether 31 (109 mg, 0;17 mmol) was dissolved in 5mL glacial acetic acid and 130 mg palladium on charcoal (10%, 0.12 mmol)was added. The mixture was stirred vigorously under a hydrogenatmosphere for 18 hours. The catalyst was removed by ultrafiltration andthe filtrate was freeze-dried to give a white foam. The latter wasdissolved in 3 mL dry acetonitrile and 92 mg tetrazole (1.31 mmol) and345 μL dibenzyl N,N-diisopropylphosphoramidite (1.03 mmol) were addedand the solution was stirred over night. Then all volatile componentswere removed in high vacuum and the residue was purified by preparativeHPLC (92% MeOH, 40 mL/min, t_(Ret)=36.5 min) to give 32 (107 mg, 67μmol, 39.4% yield) as a colorless oil.

[0384]¹H-NMR (200 MHz, CDCl₃): δ 7.45-7.09 (m, 45H, Ph), 5.26-4.79 (m,23H, CH₂-Ph, H-3, H-5, H-6), 4.65 (td, 1H, J=10.4 Hz, 3.5 Hz, H-2), 4.43(ddd, 1H, J=10.1 Hz, H-4), 2.60-2.40 (m, 1H, α-CHH(Bt)), 2.39-1.98 (m,3H, C(OPO(Bn)₂)CH₂, α-CHH(Bt)), 1.61-1.16 (m, 3H, CH), 1.02-0.82 (m, 1H,CH), 0.74 (t, 3H, ³J=7.4 Hz, CH₃), 0.72 (t, 3H, ³J=6.6 Hz, CH₃)

[0385]³¹P-NMR (81 MHz, CDCl₃): δ 0.75 (1P), 0.72 (1P), −0.01 (1P), −0.14(1P), −5.52 (1P)

[0386] MS (pos. mode, DCI, NH₃) m/z (%): 1593 [M+H⁺] (<1), 91 [Bn⁺] (18)

[0387] MS (neg. mode, DCI, NH₃) m/z (%): 1501 [M−Bn⁺] (2), 1411 [M−2Bn⁺+H⁺] (<1), 1301 [M−3 Bn⁺+2H⁺] (<1), 1131 [M−3 Bn⁺−Bt⁺+3H⁺] (2)

[0388] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-6-butyroxy-1,2,3,4,5-pentakis(phosphonooxy)-2-propyl-cyclohexane(33)

[0389] 50 mg (31 μmol) fully protected pentakisphosphate 32 wasdissolved in 5 mL glacial acetic acid, 14 mg palladium on charcoal (10%Pd, 13 μmol) was added, and the mixture was held under an hydrogenatmosphere for 12 hours. The catalyst was removed by ultrafiltration andthe filtrate was freeze-dried to give 33 (21 mg, 30 μmol, 99% yield) asa white foam.

[0390]¹H-NMR (200 MHz, DMSO-d₆): δ 5.09 (dd, 1H, ³J=9.7 Hz, J=2.0 Hz,H-6), 4.68-4.21 (m, 4H, H-2, H-3, H-4, H-5), 2.42-2.27 (m, 2H,α-CH₂(Bt)), 2.27-2.12 (m, 1H, CH(Pr)), 1.941.74 (m, 1H, CH(Pr),1.69-1.47 (m, 2H, β-CH₂(Bt), 1.35-1.11 (m, 2H, CH(Pr)), 0.87 (t, 3H,³J=7.5 Hz, CH₃(Bt)), 0.76 (t, 3H, ³J=7.5 Hz, CH₃(Pr))

[0391] 31P-NMR (81 MHz, DMSO-d₆): δ 0.74 (1P), 0.41 (1P), 0.01 (1P),−0.23 (1P), 4.9.7 (1P)

[0392] MS (pos. mode, FAB, NBA) m/z (%): 737 [M+2 Na⁺−H⁺] (1), 715[M+Na⁺] (1), 693 [M+H⁺] (<1),

[0393] MS (neg. mode, FAB, NBA) m/z (%): 713 [M+Na⁺−2H⁺] (17), 691[M−H⁺] (16), 97 [H₂PO₃ ⁻(34)

[0394] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxymethoxy)-phosphoryl]oxy]-6-butyroxy-1-propyl-cyclohexane(34)

[0395] 8 mg Pentakisphosphate 33 (12 μmol) was dissolved in 0.75 mL dryacetonitrile under an argon atmosphere and 0.1 mL diisopropylethyl amine(589 μmol) and 60 μl (600 μmol) acetoxymethyl bromide were added. Afterstirring the solution for three days in the dark all volatile componentswere removed in high vacuum and the residue was subjected to preparativeHPLC (68% MeOH, 40 mL/min, t_(Ret)=25.0 min) for purification 34 (11 mg,8 μmoles, 71% yield) was isolated as a colorless oil.

[0396]¹H-NMR (200 MHz, toluene-d₈): δ 6.02-5.53 (m, 20H, OCH₂O),5.26-4.98 (m, 3H, H-3, H-5, H-6), 4.73-4.44 (m, 2H, H-2, H-4), 2.83-2.24(m, 4H, α-CH₂(Bt), CH(Pr)), 1.68-1.23 (m, 4H, CH(Pr)), 1.97 (s, 3H,OAc), 1.93 (s, 3H, OAc), 1.86 (s, 3H, OAc), 1.86 (2s, 6H, OAc), 1.85 (s,3H, OAc), 1.84 (s, 3H, OAc), 1.83 (s, 3H, OAc), 1.80 (s, 3H, OAc), 1.78(s, 3H, OAc), 1.10 (t, 3H, ³J=7.0 Hz, CH₃(Pr)), 0.99 (s, 3H, ³J=7.3 Hz,CH₃(Bt))

[0397]³¹P-NMR (81 MHz, toluene-d₈): δ−2.89 (1P), −2.93 (1P), −3.14 (1P),−3.52 (1P), −8.70 1P)

[0398] MS (neg. mode, FAB, NBA) m/z (%): 1339 [M−CH₂OAc⁺] (11), 241[(AcOCH₂O)₂PO₂ ⁻] (100)

[0399] HR-MS: calcd. for C₄₀H₆₄O₄₀P₅ 1339.1662; found 1339.1680

EXAMPLE 5(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(12)

[0400]

[0401] Referring to the reaction scheme shown in FIG. 2, the compound(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(12) was synthesized as follows:

[0402](5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetra(benzyloxy)-5a-hydroxy-decahydro-benzo[b]oxepin(9)

[0403] 4 (0.684 g, 1.156 mmol) and dichloro tris(triphenylphosphine)ruthenium(II) (81 mg, 0.085 mmol) were suspended in 18 mL degasedmethanol under argon. The mixture was heated to reflux in the dare Afterthe solution turned homogenous 18 mg sodium borohydride (0.48 mmol) wereadded and the solution was held under reflux for another four hours.After cooling to room temperature the solution was stirred for another12 hours and finally dried under reduced pressure. The residue wasresuspended in 7.5 mL acetonitrile and 10 mL CH₂Cl₂. Then 0.14 mLtrifluoro acetic acid were added and the mixture was stirred for 5 daysat room temperature. The reaction was quenched by adding 0.3 mLdiisopropylethyl amine. All volatile components were removed underreduced pressure and the dry residue was dissolved in t-butylmethylether, extracted with phosphate buffer (pH 7) and brine. The organicphase was dried over Na₂SO₄, filtered and the solvent was evaporated.Preparative HPLC (84% MeOH, 40 mL/min, t_(Ret)=41.1 min) gave 0.433 g(63% yield) of 9 as a white solid and 0.170 g (25% yield) of thetricyclic compound 5.

[0404] m.p.: 130° C.

[0405]¹H-NMR (200 MHz, CDCl₃): δ 7.59-7.22 (m, 20H, Ph), 5.09-4.66 (m,8H, CH₂Ph), 4.10 (dd, 1H, ³J=9.6 Hz, H-9), 4.03-3.84 (m, 2H, H-2), 3.01(dd, 1H, ³J=9.1 Hz, H-7), 3.59 (d, 1H, ³J=9.6 Hz, H-8), 3.39 (d, 1H,³J=9.2, H-6), 3.29 (d, 1H, ³J=9.2 Hz, H-9a) 2.57 (s, 1H, OH), 2.41-2.24(m, 1H, oxepane), 2.13-1.59 (m, 4H, oxepane), 1.46-1.26 (m, 1H, oxepane)

[0406]¹³C-NMR (50.3 MHz, CDCl₃): δ 139.47 (C), 139.26 (C), 139.19 (C),138.62 (C), 84.25 (CH), 83.41 (CH), 83.01 (CH), 82.84 (CH), 82.27 (CH),76.78 (CH₂), 76.74 (CH₂), 76.43 (C), 76.42 (CH₂), 76.37 (CH₂), 69.79(CH₂), 41.11 (CH₂), 30.03 (CH₂), 19.74 (CH₂)

[0407] MS (pos. mode, DCI, NH₃) m/z (%): 612 [M+NH₄ ⁺] (100), 595 [M+H⁺](6), 577 [M+H⁺−H₂O] (13), 91 [Bn⁺] (77)

[0408] MS (neg. mode, DCI, NH₃) m/Z (%): 593 [M−H⁺] (2), 503 [M−Bn⁺] (5)

[0409] C₃₈H₄₂O₆ (594.75): calcd. C 76.74, H 7.12; found C 76.75, H 7.08

[0410] Synthesis of(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetra(benzyloxy)-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(10)

[0411] 9 (0.397 g, 0.669 mmol) was dissolved in 6 mL dry dimethylformamide. Sodium hydride (57 mg, 2.375 mmol) was added under vigorousstirring. After 45 min (0.130 ml, 1.148 mmol) 2-methoxy ethoxymethylchloride was added and after 12 hours additional sodium hydride (82 mg,3.417 mmol) and 2-methoxy ethoxymethyl chloride was supplied. After 90min the reaction mixture was quenched and dried in high vacuum. Theresidue was suspended in t-butylmethyl ether and washed with phosphatebuffer (pH 7) followed by brine. The organic layer was dried with Na₂SO₄and filtered. The filtrate was dried under reduced pressure und theresidue was chromatographed by preparative HPLC (88% MeOH, 40 mL/min,t_(Ret)=47.48 min) to give 0.393 g of 10 (86% yield) as a colorless oil.

[0412]¹H-NMR (200 MHz, CDCl₃): δ 7.42-7.21 (m, 20H, Ph), 5.27-5.04 (AB,2H, ²J=6.8 Hz, OCH₂O), 5.02-4.55 (m, 8H, CH₂Ph), 4.16-3.62 (m, 2H, H-2),4.02 (dd, 1H, ³J=9.5 Hz, H-7), 3.95 (dd, 1H, ³J=9.4 Hz, H-9), 3.93-3.76(m, 2H, OCH₂ CH₂O), 3.62-3.54 (m, 2H, OCH₂CH₂O), 3.51 (dd, 1H, ³J=9.2Hz, H-8), 3.40 (s, 1H, OCH₃), 3.23 (d, 1H, ³J=9.4, H-9a), 3.19 (d, 1H,³J=9.8 Hz, H-6), 2.52-2.36 (m, 1H, oxepane), 1.96-1.57 (m, 4H, oxepane),1.52-1.42 (m, 1H, oxepane).

[0413]¹³C-NMR (CDCl₃): δ 139.5 (C), 139.3 (C), 139.0 (C), 138.8(C), 91.5(CH₂), 85.1 (CH), 84.4 (CH), 84.4 (CH), 83.4 (CH), 82.3 (CH), 82.1 (C),76.6 (CH₂), 76.3 (CH₂), 76.3 (CH₂), 76.3 (CH₂), 72.3 (CH₂), 71.8 (CH₂),67.6 (CH₂), 59.4 (CH₃), 34.0 (CH₂), 31.5 (CH₂), 21.1 (CH₂)

[0414] MS (pos. mode, DCI, NH₃) m/z (%): 700 (100) [M+NH₄ ⁺], 610 (19)[M+NH₄ ⁺−Bn⁺+H⁺]593 (14) [M+2H⁺−Bn⁺], 91 (96) [Bn⁺]

[0415] MS (neg. mode, DCI, NH₃) m/z (%): 591 (14) [M−Bn⁺], 501 (15)[M−2Bn⁺+H⁺]

[0416] HR-MS: calcd. for C₃₅H₄₃O₈ [M−C₇H₇ ⁺] 591.2958, found 591.2934

[0417] Synthesis of(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetrakis[[bis(benzyloxy)phosphoryl]-oxy]-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(11)

[0418] 10 (0.381 g, 0.559 mmol) was dissolved in 5 mL ethanol and 0.05mL diisopropyl ethylamine and palladium on charcoal (10% Pd, 0.379 g,0.356 mmol) was added under an argon atmosphere at −20° C. The argonatmosphere was exchanged to hydrogen in a self-built hydrogenationapparatus and the mixture was stirred for seven days at roomtemperature. Finally, the hydrogen was exchanged to argon and thecatalyst was removed by ultrafiltration. The filtrate was dried underreduced pressure. To the foamy residue 390 mg tetrazole (5.57 mmol)dissolved in 14 mL dry acetonitrile was added. Under argon, 1.85 mL (5.5mmol) dibenzyl N,N-diisopropylphosphoramidite was added and the mixturewas stirred for 18 hours. The mixture was cooled to −40° C. and 1.2 mLperacetic acid (32% wt. 5.755 mmol) were added. The solution was allowedto warm to room temperature and the volatile components were removed inhigh vacuum. The oily residue was purified by preparative HPLC (90%MeOH, 40 mL/min, t_(R)=3 1.4 min) to give 11 (265 mg, 46% yield) as acolorless oil.

[0419]¹H-NMR (200 MHz, CDCl₃): δ 7.42-7.09 (m, 40H, Ph), 5.20-4.79 (m,20H, CH₂Ph; OCH₂O, H-7, H-9), 4.47 (q, 1H, ³J=9.8 Hz=⁴J_(PH)=9.8 Hz,H-0.8), 4.23 (dd, 1H, ³J=10.2 Hz, H-H-6), 3.82-3.53 (m, 4H, OCH₂CH₂O,OCH₂-oxepane), 3.48-3.41 (m, 2H, OCH₂CH₂O), 3.32 (s, 3H, OCH₃), 3.23((d, 1H, ³J=9.5 Hz, H-9a) 2.55-2.40 (m, 1H, oxepane), 1.91-1.87 (m, 5H,oxepane).

[0420]¹³C-NMR (50.3 MHz, CDCl₃): δ 91.2 (CH₂), 81.0 (CH), 80.5 (CH),78.5 (CH), 77.1 (CH), 75.7 (CH) 59.3 (CH₃) 33.9 (CH₂), 30.5 (CH₂), 21.2(CH₂),

[0421]³¹P-NMR (81 MHz, CDCl₃): δ 1.00 (1P), 0.61 (1P), 0.33 (1P), −0.43(1P)

[0422] MS (pos. mode, DCI, NH₃) m/z (%): 1363 (3) [M+H⁺], 91 (40) [Bn⁺]

[0423] MS (neg. mode, DCI, NH₃) m/z (%): 1271 (6) [M−Bn⁺], 277 (36)[OP(OBn)₂O]

[0424] HR-MS: calcd. for C₆₃H₇₁O₂₀P₄ [M−C₇H₇ ⁺) 1271.3489, found1271.3455

[0425] Synthesis of(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(12)

[0426] 11 (170 mg, 0.125 mmol) was dissolved in 5 mL ethanol, containing0.2 mL (1.18 mmol) diisopropylethyl amine and 319 mg Pd on charcoal (10%Pd, 3.0 mmol) suspended in ice-cold ethanol (5 mL) under argon wasadded. The argon atmosphere was exchanged to hydrogen in a self-builthydrogenation apparatus and the mixture was stirred for seven days atroom temperature. Finally, the hydrogen was exchanged to argon and thecatalyst was removed by ultrafiltration. The filtrate was dried underreduced pressure. The diisopropylethyl ammonium salt of thetetrakisphosphate was dried thoroughly and dissolved in 3 mL dryacetonitrile. Diisopropylethyl amine (0.42 mL, 2.474 mmol) andacetoxymethyl bromide (0.25 mL, 2.5 mmol) were added under argon and themixture was allowed to stir for 48 hours in the dark at roomtemperature. All volatile ingredients were removed in high vacuum andthe crude product was purified by preparative HPLC (58% MeOH, 40 mL/min,t_(Ret)=33.0) to give 12 (0.106 mg, 0.087 mmole, 70% yield) as acolorless oil.

[0427]¹H-NMR (200 MHz, d₈-toluene): δ 6.43-5.99 (m, 16H, OCH₂O),5.55-5.24 (m, 4H, H-7, H-9, OCH₂O-MEM), 5.06 (ddd, 1H, J=9.5 Hz, H-8),4.73 (dd, 1H, ³J=10.2 Hz, H-6), 4.31-3.80 (m, 6H, H-2, OCH₂CH₂O) 3.62(s, 3H, OCH₃), 3.31 (d, 1H, ³J=9.5 Hz, H-9a), 2.97-2.78 (m, 1H,oxepane), 2.39-2.24 (m, 2H, oxepane), 2.30 (s, 3H, Ac), 2.30 (s, 3H,Ac), 2.26 (s, 3H, Ac), 2.26 (s, 3H, Ac), 2.23 (s, 3H, Ac), 2.20 (s, 3H,Ac), 2.19 (s, 3H, Ac), 2.18 (s, 3H, Ac), 2.03-1.73 (m, 3H, oxepane).

[0428]¹³C-NMR (50.3 MHz, d₈-toluene): δ 169.85 (C), 169;64 (C), 169.64(C), 169.58 (C), 169.58 (C), 169.54 (C), 169.43 (C), 169.34 (C), 91.91(CH₂), 84.21-83.14 (CH₂), 81.24 (C), 80.82 (CH), 80.27 (CH), 79.34 (CH),77.87 (CH), 76.79 (CH), 72.63 (CH₂), 71.70 (CH₂), 68.71 (CH₂), 58.99(CH₃), 34.16 (CH₂), 30.87 (CH₂), 21.22 (CH₂).

[0429]³¹P-NMR (81 MHz, d₆-DMSO): δ−3.01 (1P), −3.05 (2P), −3.07 (1P)

[0430] MS (neg. mode, FAB, NBA) m/z (%): 1145 [M−CH₂OAc⁺] (7), 1073[M−2CH₂OAc⁺+H⁺] (2), 241 [(AcOCH₂O)₂PO₂] (100)

[0431] MS (pos. mode, FAB, NBA) m/z (%): 1240 [M+Na⁺] (4), 1147[M+2H⁺−CH₂OAc⁺] (12)

[0432] HR-MS: calcd. for C₃₅H₅₇O₃₄P₄ [M−CH₂OAc⁺] 1145.1672, found1145.1696

EXAMPLE 6 Synthesis of (4aα, 5α,6β,7α,8β,8aα)-5,6,7,8-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-4a-butyryloxy-octahydro-chromen(19)

[0433]

[0434] Referring to the reaction scheme shown in FIG. 3, the compound(4aα,5α,6β,7α,8β,8aα)-5,6,7,8-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-4a-butyryloxy-octahydro-chromen(19) was synthesized as follows:

[0435] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-6-allyloxy-2,3,4,5-tetra-benzyloxy-1-hydroxy-1-vinyl cyclohexane(13)

[0436] 341 mg 2 (0.59 mmol) were suspended in 20 mL absolute diethylether and the mixture was cooled to 40° C. Under an argon atmosphere0.65 mL (1.12 mmol) of a 1.7 M solution of vinylmagnesium chloride inTHF was added in two equal portions. The solution was stirred for 18hours at room temperature, when the reaction was quenched by adding 10mL phosphate buffer (0.5 M, pH 7).

[0437] The organic layer was diluted with t-butylmethyl ether, washedtwice with phosphate buffer and brine, dried over anhydrous sodiumsulfate, and filtered. The volatile components were removed underreduced pressure. From the oily residue the two 2-hydoxy epimers werepurified by preparative HPLC (RP-18, 40 mL/min, 88% MeOH, t_(Ret)=32.05min and 36.40 min, respectively) to give epimer 13 with the axialhydroxy group in 29% (0.104 g, 0.17 mmol) as a colorless solid, andepimer epi-13 with the equatorial hydroxy group in 38% yield (0.136 g,0.23 mmol) as colorless oil, respectively.

[0438] m.p. 13: 110.5° C.

[0439]¹H-NMR (200 MHz, CDCl₃): δ 7.41-7.21 (m, 20H, Ph), 6.02-5.72 (m,2H, allyl), 5.56 (dd, 1H, ²J=1.5 Hz ³J=17.0 Hz, allyl), 5.39 (dd, 1H,²J=1.4 Hz ³J=10.1 Hz, allyl), 5.28-5.10 (m, 2H, allyl), 4.974.84 (m, 6H,OCH₂Ph), 4.844.60 (AB, 2H, OCH₂Ph), 4.34-4.07 (m, 2H, allyl), 3.98 (dd,1H, ³J=3.3 Hz, 9.5 Hz, H-5), 3.95 (dd, 1H, ³J=9.3 Hz, H-3), 3.54(dd, 1H,³J=9.5 Hz, H-4), 3.39 (d, 1H, ³J=9.3 Hz, H-6), 3.27 (d, 1H, ³J=9.3 Hz,H-2), 2.58 (s, 1H, OH)

[0440]¹³C-NMR (50.3 MHz, CDCl₃): δ 75.4 (1C), 76.4 (2C), 76.4 (1C), 76.5(2C), 82.1 (1C), 82.5 (1C), 82.9 (1C), 83.0 (1C), 83.3 (1C), 117.1 (1C),117.7 (1C), 128.1, 128.1, 128.3, 128.3, 128.5, 128.7, 128.8, 135.4 (1C),140.4 (1C)

[0441] MS (pos. mode, FAB, NBA) m/z (%): 624 [M+NH₄ ⁺] (100), 607 [M+H⁺](<1), 534 [M+NH₄ ⁺+H⁺−Bn⁺] (5), 515 [M−Bn⁺] (11), 425 M−2 Bn⁺+H⁺] (2),335 [M−3 Bn⁺+2H⁺] (1)

[0442] MS (neg. mode, FAB, NBA) m/z (%): 605 [M−H⁺]] (63), 565 [M−C₃H₅⁺] (13), 515 [M−Bn⁺] (100), 425 [M−2 Bn⁺+H⁺] (10)

[0443] C₃₉H₄₂O₆ (606.76): calcd. C 77.20, H 6.98; found C 77.21, H 7.04

[0444] Synthesis of (4aα, 5α,6β,7α,8β,8aα)-5,6,7,8-tetra(benzyloxy)-4a-hydroxy-4a,5,6,7,8,8a-hexahydro-[2H]-chromen(14)

[0445] 270 mg 13 (0.446 mmol) were dissolved in 50 mL dry methylenechloride. Bis(triccylohexylphosphin)benzylidene ruthenium (IV)dichloride (Grubb's catalyst, 5.8 mg, 7 μmol) was added under argon andthe solution was stirred for 24 hours in the dark. Then another 7.9 mg(10 mmol) Grubb's catalyst was added and the mixture was heated toreflux for 90′ min. After reaching room temperature methylene chloridewas added and the organic layer was extracted with phosphate buffer (0.5M, pH 7) and brine. Then it was dried, over anhydrous sodium sulfate,filtered, and the solvent was removed under reduced pressure. The dryresidue was chromatographed by preparative HPLC (RP18, 90% MeOH, 40mL/min, t_(Ret)=20.5 min) to give 14 in 93% yield (237.8 mg, 0.41 mmol)as a colorless solid.

[0446] m.p.: 128.5° C.

[0447]¹H-NMR (200 MHz, CDCl₃): δ 7.50-7.27 (m, 20H, Ph), 6.11 (dt, 1H,³J=10.1 Hz, ⁴J=2.0 Hz, H-4), 5.94 (ddd, 1H, ³J=10.1 Hz, ⁴J=3.1 Hz,⁴J=1.6, H-3), 5.134.66 (m, 8H, OCH₂Ph), 4.49-4.21 (m, 2H, H-2), 4.21(dd, 1H, ³J=9.3 Hz, H-8), 4.07 (dd, 1H, ³J=9.3 Hz, H-6), 3.64 (dd, 1H,³J=9.1 Hz, H-7), 3.38 (d, 1H, ³J=7.8 Hz, H-5), 3.33 (d, 1H, ³J=7.4 Hz,H-8a), 2.23 (s, 1H, OH)

[0448]¹³C-NMR (50.3 MHz, d₆-DMSO): δ 140.00 (C), 139.76 (C), 139.65 (C),139.41 (C), 129.81, 127.40, 83.89 (CH), 83.83 (CH), 82.23 (CH), 81.19(CH), 80.18 (CH), 75.97 (CH₂), 75.67 (CH₂), 75.55 (CH₂), 74.75 (CH₂),68.30 (C), 66.62 (CH₂)

[0449] MS (neg. mode, FAB, NBA) m/z (%): 731 [M+NBA] (<1), 577 [M−H⁺](<1)

[0450] MS (pos. Mode, FAB, NBA) m/z (%): 579 [M+H⁺] (3), 91 [Bn⁺] (100)

[0451] C₃₇H₃₈O₆ (578.71): calcd C 76.79, H 6.62; found C 76.61, H 6.54

[0452] Synthesis of (4aα,5α,6β,7α,8β,8aα)-5,6,7,8-tetra(benzyloxy)-4a-hydroxy-octahydro-chromen(15)

[0453] 14 (0.238 g, 0.412 mmol) was dissolved in 10 mL toluene/acetone(1:1, v/v) and 78 mg (84 μmol) chlorotris(triphenylphosphine)rhodium(I)(Wilkinson's catalyst) were added. The mixture was subjected to ahydrogen atmosphere in a self-built hydrogenation apparatus and stirredin the dark for seven days, during which time the solution turned fromyellow to deep red. The reaction was stopped by flooding the apparatus'with argon. The solvents were removed under reduced pressure and theresidue was dissolved in 100 mL t-butylmethyl ether and washed twicewith brine. The organic phase was dried over anhydrous sodium sulfateand filtered. After evaporation the residue was purified by preparativeHPLC (RP18, 40 mL/min, 88% MeOH, t_(Ret)=33.00 min) to give 15 as aslightly yellow oil (0.124 g, 52% yield).

[0454]¹H-NMR (200 MHz, CDCl₃): δ 7.44-7.21 (m, 20H, Ph), 5.05-4.59 (m,8H, OCH₂Ph), 4.22-4.08 (m, 1H, H-2), 4.05 (dd, 1H, ³J=9.4 Hz, H-6), 3.91(dd, 1H, ³J=9.34 Hz, H-0.8), 3.56 (dd, 1H, ³J=9.34 Hz, H-7), 3.53-3.39(m, 1H, H-2) 3.24 (d, 1H, ³J=9.7 Hz, H-5), 3.22 (d, 1H, ³J=9.7 Hz,H-8a), 2.16-1.95 (m, 2H, tetrahydropyrane(thp)-ring), 1.57-1.26 (m, 2H,thp-ring)

[0455] MS (neg. mode, DCI, NH₃) m/z (%): 579 [M−H⁺] (70), 489 [M−Bn⁺](43),

[0456] MS (EI): 489 [M−Bn⁺] (42), 91 (Bn⁺) (100)

[0457] HR-MS: calcd. for C₃₀H₃₃O₆ [M−Bn⁺] 489.2284, found 489.2260

[0458] Synthesis of (4aα,5α,6β,7α,8β,8aα)-5,6,7,8-tetra(benzyloxy)-4a-butyryloxy-octahydro-chromen(16)

[0459] 15 (0.120 g, 0.207 mmol) was dissolved in 7.5 mL dry toluene.After adding 0.1 mL (5.733 mmol) butyric acid chloride and 0.35 mL(1.376 mmol) phosphazen base-P₁-t-Bu the mixture was heated to refluxfor 12 hours. The mixture was dried in high vacuum, the residue wasdissolved in t-butylmethyl ether and the organic phase was extractedwith phosphate buffer (0.5 mM, pH 7) and brine. The solution was driedover anhydrous sodium sulfate, filtered, and the solvent was evaporatedunder reduced pressure. The residue was purified by preparative HPLC(RP18, 40 mL/min, 90% MeOH, t_(Ret)=44.13 min). 16 was a colorless oil(0.078 g, 58% yield).

[0460]¹H-NMR (200 MHz, CDCl₃): δ 7.45-7.18 (m, 20H, Ph), 5.00-4.70 (m,8H, OCH₂Ph), 4.12 (dd, 1H, ²J=11.3 Hz, ³J=5.0 Hz, H-2 eq), 3.91 (dd, 1H,³J=9.3 Hz, H-8), 3.89-3.75 (m, 1H, H-eq), 3.82 (dd, 1H, ³J=9.5 Hz, H-6),3.62-3.43 (m, 1H, H-2ax), 3.55 (dd, 1H, ³J=9.2 Hz, H-7), 3.31 (d, 1H,³J=9.4 Hz, H-5), 3.15 (d, 1H, ³J=9.6 Hz, H-8a), 2.53-2.27 (m, 2H,α-CH₂(Bt)), 1.95-1.79 (m, 1H, thp-ring), 1.88-1.66 (m, 2H, β-CH₂ (Bt)),1.66-1.53 (m, 1H, thp-ring), 1.38-1.19 (m, 1H, thp-ring), 1.01(t, 3H,³J=7.4 Hz, CH₃)

[0461]¹³C-NMR (50.3 Mhz, CDCl₃): δ=172.7 (C), 139.3 (C), 138.9 (C),138.8 (C), 138,7 (C), 128.9, 128.8, 128.8, 128.7, 128.6, 128.5, 128.5,128.2, 128.1, 128.0, 128.0, 84.2 (CH), 83.5 (CH), 83.0 (CH), 83.0 (CH),82.66 (C), 81.1 (CH), 76.7(CH₂), 76.6 (CH₂), 76.5 (CH₂), 76.1 (CH₂),68.6 (CH₂), 38.4 (CH₂), 30.2 (CH₂), 22.4 (CH₂), 19.1 (CH₂), 14.2 (CH₃)

[0462] MS (neg. mode, DCI, NH₃): m/z (%): 649 (5) [M−H⁺], 579 (61)[M−H⁺−Bt], 559 (100) [M−C₇H₇ ⁺], 489 (18) [M−C₇H₇ ⁺−Bt], 469 (20)[M−C₇H₇ ⁺−C₇H₇]

[0463] HR-MS: calcd. for C₃₄H₃₉O₇ [M−Bn⁺] 1559.2696; found 559.2720

[0464] Synthesis of(4aα,5α,6β,7α,8β,8aα)-5,6,7,8-tetrakis[[bis(benzyloxy)phosphoryl]oxy]-4a-butyryloxy-octahydro-chromen(17)

[0465] 75.4 mg (0.116 mmol) 16 and palladium on charcoal (10% Pd, 149mg, 0.140 mmol) were suspended in 10 mL ethanol under argon and at −20°C. The atmosphere was exchanged to hydrogen in a self-builthydrogenation apparatus and stirred for 10 days. The mixture wasultrafiltered and the filtrate was dried under reduced pressure.

[0466] The residue and tetrazole (85.1 mg, 1.22 mmol) were dissolved in3 mL dry acetonitrile under argon.Dibenzyl-N,N-diisopropylphosphoramidite (390 μl, 1.16 mmole) was addedand the mixture was stirred for 18 hours at room temperature.

[0467] The flask was cooled to −40° C. and 0.285 mL (1.356 mmole)peracetic acid (32% by weight) was added. The solution was allowed towarm to room temperature and the volatile components were removed inhigh vacuum. Crude 17 was purified by preparative HPLC (RP18, 88% MeOH,40 mL/min, t_(ret)=50.25 min) to give a clear oil, that containedsubstantial amounts of a by-product 17a that where the 4a-O-butyratemigrated to the 5-hydroxy group. Both compounds were separated by asecond preparative HPLC run (ChiraDex, 40 mL/min, 100% MeCN).4a-phosphorylated by-product 17a eluted with t_(ret)=31.0 min. (15 mg,11.3 μmol, 10% yield), while the desired compound 17 had a t_(Ret)=120min (16.8 mg, 12.6 μmol, 11% yield).

[0468]¹H-NMR (200 MHz, CDCl₃): δ 7.46-7.06 (m, 20H, Ph), 5.17-4.58 (m,19H, CH₂-Bn, H-6, H-7, H-8), 4.43 (dd, 1H, ³J_(HH)=³J_(PH)=9.1 Hz, H-5),4.00-3.27 (m, 1H, thp-ring), 3.81-3.68 (m, 1H, thp-ring), 3.42-3.21 (m,1H, thp-ring), 3.30 (d, 1H, ³J=9.7 Hz, H-8a), 2.47-2.20 (m, 2H,α-CH₂(Bt)), 1.81-1.49 (m, 5H, thp, β-CH₂ (Bt)), 0.93 (t, 3H, ³J=7.4,CH₃)

[0469]¹³C-NMR (50.3 MHz, CDCl₃): δ 172.04 (C), 136.88-136.15 (C-Ph),129.02-127.89 (CH-Ph), 80.47 (C), 79.62 (CH), 78.66 (CH), 77.21 (CH),76.84 (CH), 75.35 (CH), 70.45-69.41 (CH₂-Bn), (CH₂), 68.49 (CH₂), 37.72(CH₂), 30.16 (CH₂), 21.66 (CH₂), 18.97 (CH₂), 14.10 (CH₃)

[0470]³¹P-NMR (81 MHz, CDCl₃): δ 0.40 (1P), 0.30 (1P), 0.15 (1P), −0.97(1P)

[0471] MS (neg. mode, FAB, NBA) m/z (%): 1329 [M−H⁺] (<1), 1239 [M−Bn⁺](5)

[0472] HR-MS: calcd. for C₆₂H₆₇O₁₉P₄ [M−Bn⁺] 1239.3228, found 1239.3210

[0473] (4aα, 5α,6β,7α,8β,8aα)-4a,6,7,8-Tetrakis[[bis(benzyloxy)phosphoryl]oxy]-5-butyryloxy-octahydro-chromen(17a)

[0474]¹H-NMR (200 MHz, CDCl₃): δ 7.53-7.07 (m, 40H, Ph), 5.39-4.68 (m,20, CH₂Ph, H-5, H-6, H-7, H-8), 3.83 (dd, 1H, ²J=10.9 Hz, ³J 4.2 Hz, H-2eq), 3.45 (dd, 1H, ³J=9.7 Hz, ⁴J_(PH)=5.9 Hz, H-8a), 3.35 (m, 1H,thp-ring), 2.96 (m, 1H, thp-ring), 2.17-1.89 (m, 1H, thp-ring),1.76-1.47 (m, 4H, thp-ring, β-CH₂ (Bt)), 0.86 (t, 3H, ³J=7.4 Hz, CH₃)

[0475]³¹C-NMR (50.3 MHz, CDCl₃>: δ 173.75 (C), 136.88-136.15 (C),129.20-127.51 (CH), 81.84 (C), 79.07 (CH), 77.95 (CH), 76.97 (CH), 76.97(CH), 72.97 (CH), 70.03-69.33 (CH₂-Bn), 68.49 (CH₂), 35.97 (CH₂), 29.63(CH₂), 20.96 (CH₂), 18.12 (CH₂), 14.04 (CH₃)

[0476] 31P-NMR (81 MHz, CDCl₃): 8-0.02 (1P), −0.45 (1P), −0.86 (1P),−5.28 (1P)

[0477] MS (pos. mode, DCI, NH₃) m/z (%): 1331 [M+H⁺] (2), 1241[M+2*H⁺−Bn⁺] (3)

[0478] MS (neg. mode, DCI, NH₃) m/z (%): 1329 [M−H⁺] (<1), 1239 [M−Bn⁺](9), 1149. [M+H⁺−2*Bn⁺] (4)

[0479] HR-MS calcd. for C₆₂H₆₇O₁₉P₄ [M−Bn⁺] 1239.3228, found 1239.323

[0480] Synthesis of (4aα,5α,6β,7α,8β,8aα)-4a-butyryloxy-5,6,7,8-tetrakis(phosphonooxy)-octahydro-chromen(18)

[0481] 17 (6.7 mg, 12.6 μmol) was dissolved in 5.5 mL glacial aceticacid and 185 mg palladium on charcoal (10% Pd, 174 μmol) was added. Themixture was stirred under a hydrogen atmosphere for 12 hours. Then thecatalyst was removed by ultrafiltration (0.45 μm RC55 membrane filter)and the filtrate was freeze-dried to yield 7.5 mg (99% yield) of 18 as acolorless oil.

[0482]¹H-NMR (200 MHz, d₆-DMSO): δ 4.44 (ddd, 1H, J=9.6 Hz, H-8),4.314.02 (m, 3H, H-6, H-7, H-2), 3.88 (d, 1H, J=9.5 Hz, H-5), 3.52-3.21(m, 3H, H-8a, H-2, thp-ring), 2.23 (t, 2H, J=7.2 Hz, α-CH₂ (Bt)),1.62-1.29 (m, 7H, β-CH₂ t), thp-ring), 0.89 (t, 3H, J=7.2 Hz)

[0483]¹³C-NMR (50.3 MHz, d₆-DMSO): δ 171.43 (C), 82.08, 67.92 (CH₂),δ8.16 (CH₂), 29.08 (CH₂), 21.96 (CH₂), 18.95 (CH₂), 14.41 (CH₃)

[0484]³¹P-NMR (81 MHz, d₆-DMSO): δ 0.89 (2P), 0.16 (1P), −0.18 (P)

[0485] MS (neg. mode, FAB, glycerol) m/z (%): 631 [M−2H⁺+Na⁺] (5), 609[M−H⁺] (26)

[0486] Synthesis of (4aα, 5α,6β,7α,8β,8aα)-5,6,7,8-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-4a-butyryloxy-octahydro-chromen(19)

[0487] 18 (7.2 mg, 11.8 μmol) was dissolved in 0.7 mL dry acetonitrileand diisopropylethyl amine (62 μl, 365 μmol) and acetoxymethyl bromide(36 μL, 365 μmol) were added under an argon atmosphere. After stirring36 hours at room temperature all volatile components were removed inhigh vacuum and the residue was extracted several times with 1 mL drytoluene. The combined extracts were centrifuged and subsequently thesolvent was removed under reduced pressure to give 19 (8 mg, 6 μmol, 50%yield) as a colorless oil.

[0488]¹H-NMR(200 MHz, d₈-toluene): δ 6.01-5.62 (m, 16H, OCH₂O),5.07-4.52 (m, 4H, H-5, H-6, H-7, H-8), 3.92-3.78 (m, 1H, thp-ring),3.78-3.65 (m, 1H, H-2eq), 3.27-3.12 (m, 1H, H-2ax), 2.91 (d, 1H, ³J=9.2Hz, H-8a), 2.31-2.13 (m, 2H, α-CH₂(Bt)), 1.89 (s, 3H, OCH₃), 1.88 (s,3H, OCH₃), 1.85 (s, 3H, OCH₃), 1.84 (s, 3H, OCH₃), 1.81 (s, 3H, OCH₃),1.76 (s, 3H, OCH₃), 1.75 (s, 3H, OCH₃), 1.74 (s, 3H, OCH₃), 1.69-1.30(m, 5H, P—CH₂, thp-ring), 0.88 (t, 3H, ³J=7.4 Hz, CH₃(Bt))

[0489]¹³C-NMR (50.3 MHz, d₈-toluene): δ 83.65-82.94 (OCH₂O), 78.70 (CH),78.28 (CH), 77.59 (CH), 77.25 (CH), 76.08 (CH), 67.74 (CH₂), 37.16(CH₂), 28.94 (CH₂), 21.72 (CH₂). 21.62-19.83 (CH₃), 18.83 (CH₂), 13.72(CH₃)

[0490]³¹P-NMR (81 MHz, d₈-toluene): δ −2.98 (1P), −3.14 (1P), −3.29(1P), −3.36 (1P)

[0491] MS (neg. mode, FAB, NBA) m/z (%): 1113 [M−CH₂OAc⁺] (9), 1041′[M−2 CH₂OAc⁺+H⁺] (5), 969 [M−3 CH₂OAc⁺+2H⁺] (2), 241 [(AcOCH₂O)₂PO₂ ⁻](94)

EXAMPLE 7 Synthesis of(1S 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.0^(1,6)]-dodecane (8)

[0492]

[0493] Referring to the reaction scheme shown in FIG. 1, the compound(1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.0^(1,6)]-dodecane(8) was synthesized as follows:

[0494] Synthesis of 1-O-allyl-3,4,5,6-tetra-O-benzyl-myo-inosose (2)

[0495] 1.44 g (2.5 mmol) 1-O-Allyl-3,4,5,6-tetra-O-benzyl-myo-inositol 1was dissolved in 10 mL DMSO and 1 mL acetic acid anhydride (10.6 mmol)was added under an argon atmosphere. After 48 hours at room temperatureall volatile ingredients were removed in high vacuum and the crudeyellow residue was recrystallized twice from methanol to give 1.13 g(2.0 mmol, 79% yield) of 2.

[0496] m.p.: 124.3° C.

[0497]¹H-NMR (200 MHz, CDCl₃): δ 7.53-7.23 (m, 20H, Ph), 6.14-5.90 (m,H,OCH₂CHCH₂), 5.47-5.20 (m, 2H, OCH₂CHCH₂), 5.06-4.57 (m, 8H, OCH₂Ph),4.49-4.34 (m, 1H, OCHHCHCH₂), 4.24 (dd, 1H, ³J=9.9 Hz, 1.6 Hz, H-1),4.21-4.07 (m, 1H, OCHHCHCH₂), 4.16 (dd, 1H, ³J=9.9 Hz, 1.6 Hz, H-3)33.95(dd, 1H, H-5, ³J=9.1 Hz, H-5), 3.69 (dd, 1H, H-4, ³J=9.7 Hz, H-4), 3.66(dd, 1H, ³J=9.6 Hz, H-6)

[0498]¹³C-NMR (50.3 MHz, d₆DMSO): δ 203.44 (C), 139.37 (C), 139.37 (C),39.31 (C), 139.14 (C), 135.83 (CH), 129.18-128.21 (CH, Bn) 117,25 (CH₂),84.46 (CH), 84.30 (CH), 82.15 (CH), 81.57 (CH), 81.51 (CH), 75.79 (CH₂),75.50 (CH₂), 75.50 (CH₂), 73.03 (CH₂), 72.16 (CH₂)

[0499] MS (pos. mode, NH₃, DCI) m/z (%): 579 [M+H⁺] (10), 489[M+2*H⁺−Bn⁺] (68), 399 [M+3*H⁺−2*Bn⁺] (6), 108 [Bn⁺+NH₃] (100)

[0500] C₃₇H₃₈O₆ (578.69): calcd. C 76.79, H 6.61; found C 76.54, H 6.53

[0501] Synthesis of rac-(1S, 2R, 3S, 4R, 5S,6S)-1-allyl-6-allyloxy-2,3,4,5-tetra-benzyloxy- 1-hydroxy-cyclohexane(3)

[0502] 2 (561 mg, 0.97 mmol) was suspended in 10 mL absolute diethyl andthe mixture was cooled to −25° C. 0.55 mL (1.10 mmol) of a 2-molarsolution of allyl magnesium chloride in THF was added under argon andvigorous stirring. After 20 min another 0.1 mL (0.2 mmol) allylmagnesium chloride solution was added. The mixture was allowed to warmup and was quenched with 10 mL 0.5 M phosphate buffer, pH 7, after 14hours. The organic layer was diluted with t-butylmethyl ether, extractedwith brine and dried over anhydrous sodium sulfate. After filtration andevaporation of the solvent in vacuum the solid residue waschromatographed by preparative HPLC (RP-18, 40 mL/min, 90% MeOH,t_(Ret)=34.50 min) to give 377 mg 3 as a colorless solid (0.61 mmol, 63%yield).

[0503] m.p.: 97.5° C.

[0504]¹H-NMR (200 MHz, CDCl₃): δ 7.53-7.32 (m, 20H, Ph), 6.19-5.95 (m ,1H, OCH₂CHCH₂), 5.92-5.68 (m, 1H, HOCCH₂CH═CH₂), 5.42-4.72 (m, 8H,CH₂Ph), 4.48 (dd, 1H, ²J=12.0 Hz, ³J=5.7 Hz, OCHHCH═CH₂), 4.15 (dd, 1H,²J=12.1 Hz, ³J=5.7 Hz OCHHCH═CH₂), 4.03 (dd, 1H, ³J=9.4 Hz, H-3), 3.98(dd, 1H, ³J=9.4 Hz, H-5), 3.48 (dd, 1H, ³J=9.8 Hz, H₄), 3.38 (d, 1H,³J=9.5 Hz, H-6), 3.25 (d, 1H, ³J=9.4 Hz, H-2), 2.61 (d, 1H, HOCCH₂CHCH₂,³J=7.2 Hz), 2.31 (br s, 1H, OH)

[0505]¹³C-NMR (50.3 MHz, CDCl₃): δ 139.08 (C), 139.08 (C), 138.96 (C),138.65 (C), 135.34 (CH), 133.33 (CH), 129.18-127.51 (CH, Bn), 119.92(CH₂), 117.39 (CH₂), 83.69 (CH), 83.55 (CH), 83.55 (CH), 80.43 (CH),80.17 (CH), 78.02 (C), 76.31 (CH₂), 76.31 (CH₂), 76.31 (CH₂), 75.87(CH₂), 75.17 (CH₂), 39.94 (CH₂)

[0506] MS (pos. mode, FAB, NBA) m/z (%): 621 [M+H⁺] (3), 91 [Bn⁺] (100)

[0507] MS (neg. mode, FAB, NBA) m/z (%): 772 [M+NBA-H⁺] (1), 619 [M−H⁺](1), 503 [M−BnO⁺] (14)

[0508] C₄₀H₄₄O₆ (620.79): calcd. C 77.39, H 7.14; found C 77.21, H 7.04

[0509] Synthesis of(5aα,6α,7β,8α,9β,9aα)-6,7,8,9-tetrabenzyloxy-5a-hydroxy-2,5,5a,6,7,8,9,9a-octahydro-benzo[b]oxepin(4)

[0510] 3 (353 mg, 0.57 mmol) was dissolved in 50 mL dry methylenechloride. 5.2 mg (6.3 μmol)bis(tricyclohexylphosphin)benzylidene-ruthenium (IV)-dichloride wasadded under an argon atmosphere, and the solution was stirred for 72hours in the dark. The organic layer was diluted with methylene chlorideand washed several times with phosphate buffer pH 7 and brine. Afterdrying over sodium sulfate and filtration, the organic layer was driedand the crude residue was purified by preparative HPLC (RP-18, 40mL/min, 90% MeOH, t_(Ret)=25.25 min) to give 318 mg (0.54 mmol, 94%yield) 4 as a colorless solid.

[0511] m.p.: 123.0° C.

[0512]¹H-NMR (200 MHz, CDCl₃): δ 7.48-7.23 (20H, Ph), 6.09-5.95 (m, 1H,H-3), 5.80-5.65 (m, 1H, H-4), 5.17-4.67 (m, 8H, OCH₂Ph), 4.50 (dd,1H¹²J=15.4 Hz, ³J=5.6 Hz, H-2), (4.25-4.07, m, 1H, H-2), 4.12 (dd, 1H,³J=9.6 Hz, H-7), 3.87 (dd, 1H, ³J=9.2 Hz, H-9), 3.60 (dd, 1H, ³J=9.5 Hz,H-8), 3.51 (d, 1H, ³J=9.2 Hz, H-9a), 3.27 (d, 1H, ³J=9.7 Hz, H-6), 2.97(dd, 1H, ²J=15.8 Hz, ³J=7.8 Hz, H-5), 2.64 (s, 1H, OH), 2.22-2.07 (m,1H, H-5)

[0513]³C-NMR (50.3 MHz, CDCl₃): δ 139.46 (C), 139.16 (C), 139.02 (C),138.35 (C), 132.33 (CH), 129.00-127.95 (CH, Bn),128.33 (CH), 88.53 (CH),83.36 (CH), 82.79 (CH), 82.74 (CH), 82.36 (CH), 76.64 (CH₂), 76.64(CH₂), 76.34 (CH₂), 76.30 (CH₂), 74.29 (C), 69.72 (CH₂), 37.42 (CH₂)

[0514] MS pos. mode, FAB, NBA) m/z (%): 591 [M−H⁺] (1), 91 [Bn⁺] (100)

[0515] MS (neg. mode, FAB, NBA) m/z (%): 745 [M+NBA⁻] (2), 591 [M−H⁺](<1)

[0516] C₃₈H₄₀O₆ (592.73): calcd. C 77.00, H 6.80; found C77.03, H 6.73

[0517] Synthesis of(1S,2R,3S,4R,5S,6S,8R)-2,3,4,5-tetrabenzyloxy-7,12-dioxa-tricyclo-[6.3.1.0^(1,8)]-dodecane(5)

[0518] 4 (0.138 g, 0.233 mmol) was dissolved in 10 mL aqueous ethanol(90%). Tris(triphenylphosphin)rhodium-(I)-chloride (46 mg, 0.43 mmol)and 0.04 mL diisopropylethyl amine (0.234 mmole) were added under argon.The mixture was heated to reflux in the dark for 18 hours. Under reducedpressure the mixture was dried, the residue dissolved in t-butylmethylether and washed with phosphate buffer pH 7 and brine. The organic layerwas dried over sodium sulfate and filtered. The filtrate was dried andthe crude residue was prepurified by preparative HPLC (RP18, 90% MeOH,40 mL/min). The resulting mixture was dissolved in 5 mL chloroform, 0.1mL trifluoro acetic acid was added, and the mixture was stirred for 3days at room temperature. The solution was diluted with methylenechloride and extracted with phosphate buffer pH 7 and brine. The organiclayer was dried over sodium sulfate and filtered. The filtrate was driedand the crude residue was purified by preparative HPLC (RP18, 90% MeOH,40 mL/min, t_(Ret)=32.10 min). 5 was isolated as a colorless solid(0.050g, 0.084 mmol, 36% yield). As a by-product 0.014 g (0.026 mmol,10% yield) 6,7,8,9-tetrabenzyloxy-5a-hydroxydecahydrobenzo-oxepin wasisolated.

[0519] m.p.: 106.3° C.

[0520]¹H-NMR (200 MHz, CDCl₃): δ 7.60-7.25 (20H, Ph), 5.73 (br s, 1H,OCHO), 5.16-4.75 (m, 8H, OCH₂Ph), 4.18 (d, 1H, ³J=6.1 Hz, H-6), 4.00(dd, 1H, ³J=9.5 Hz, H-3), 3.71 (dd, 1H, ³J=9.8 Hz, 6.2 Hz, H-5), 3.53(dd, 1H, ³J=9.5 Hz, H-4), 3.52 (d, 1H, ³J=9.7 Hz, H-2), 2.36-1.64 (m,5H, thp-ring), 1.40-1.22 (m, 1H, thp-ring)

[0521]¹³C-NMR (50.3 MHz, CDCl₃): δ 139.14 (C), 139.14 (C), 139.01 (C),138.41 (C), 103.87 (CH), 86.08 (CH), 83.20 (CH), 83.10 (C), 82.53 (CH),81.72 (CH), 81.67 (CH), 76.90 (CH₂), 76.18 (CH₂), 76.00 (CH₂), 74.62(CH₂), 31.27 (CH₂), 30.60 (CH₂), 16.59 (CH₂)

[0522] MS (neg. mode, DCI, NH₃) m/z (%): 591 [M−H⁺], 501 [M−Bn⁺], 411,[M−2Bn⁺+H⁺], 321 [M−3Bn⁺+2H⁺]

[0523] MS (pos. mode, DCI, NH₃) m/z (%): 610 [M+NH₄ ⁺], 593 [M+H⁺], 520[M+NH₄ ⁺−Bn]

[0524] C₃₈H₄₀O₆ (592.73): calcd. C77.00, H 6.80; found C 77.19, H 6.69

[0525] Synthesis of (1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis[[bis(benzyloxy)-phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.01,6]-dodecane(6)

[0526] 5 (0.159 g, 0.267 mmol) was dissolved in 5.5 mL ethanol at −20°C. A suspension of 0.222 g palladium on charcoal (10% Pd, 0.209 mmol) in5.5. mL ethanol under an argon atmosphere at −20° C. was added. Theargon atmosphere was exchanged to hydrogen in a self-built hydrogenationapparatus and the mixture was stirred for eight days at roomtemperature. Finally, the hydrogen was exchanged to argon and thecatalyst was removed by ultrafiltration The filtrate was dried in highvacuum and the residue was dissolved in dry acetonitrile. Then tetrazole(160 mg, 2.286 mmol) and dibenzyl diisopropylphosphoramidite (0.705 ml,2.098 mmol) were added under argon and the mixture was stirred for 18hours. After cooling to −0° C., peracetic acid (32% in acetic acid, 0.44mL, 2.1 mmol) was added. The solution was allowed to warm to roomtemperature and all volatile components were removed in high vacuum. Theoily residue was chromatographed by preparative HPLC (88% MeOH,. 40mL/min, t_(Ret)=38.45 min) to give 6 (0.173 mg, 0.136 mmol, 51% yieldfrom 5) as a colorless oil.

[0527]¹H-NMR (200 MHz, CDCl₃): δ 7.45-7.17 (m, 8H, Ph), 5.50 (br s, 1H,OCHO), 5.19-4.77 (m, 20H, CH₂Ph, H-2, H-3, H-4, H-5), 4.18 (d, 1H,³J=4.7 Hz, H-6), 2.12-1.90 (m, 1H, thp-ring), 1.82-1.45 (m, 5H,thp-ring)

[0528]³¹P-NMR (81 MHz, CDCl₃): 8−0.16 (1P), −0.35 (1P), −0.95 (2P)

[0529]¹³C-NMR (50.3 MHz, CDCl₃): δ 136.18 (C), 136.17 (C), 136.17 (C),136.16 (C), 104.23 (CH), 81.22 (C), 78.77 (CH), 78.20 (CH), 77.76 (CH),77.31 (CH), 31.33 (CH₂), 30.22 (CH₂), 16.17 (CH₂)

[0530] MS (neg. mode, DCI, NH₃) m/z (%): 1271 [M−H+], 1181 [M−Bn⁺],1091, [M−2Bn⁺+H⁺]

[0531] MS (pos. mode, DCI, NH₃) m/z 12906[M+NH₄ ⁺], 1272 [M+H⁺]

[0532] HR-MS: calcd. for C₅₉H₆₁O₁₈P₄ [M−Bn⁺] 1181.2809, found 1181.2819

[0533] Synthesis of(1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis(phosphonooxy)-7,12-dioxa-tricyclo-[6.3.1.0^(1,6)]-dodecane(7)

[0534] Under an argon atmosphere 6 (168 mg, 0.132 mmol) was dissolved in3 mL ethanol cooled to −20° C. A suspension of 190 mg palladium oncharcoal (10% Pd, 0.179 mmol) in 5 mL ethanol under an argon atmosphereat −20° C. was added. The argon atmosphere was exchanged to hydrogen ina self-built hydrogenation apparatus and the mixture was stirred forseven days at room temperature. Finally, the hydrogen was exchanged toargon and the catalyst was removed by ultrafiltration. The filtrate wasdried in high vacuum to give 7 as the free acid (60 mg, 0.108 mmol). 33mg were treated with Dowex 50 WX8, sodium form to give 7 as the sodiumsalt (17 mg, 0.023 mmole, 17% yield).

[0535]¹H-NMR (200 MHz, D₂O): δ 5.63 (br s, 1H, OCH₂O), 4.32-3.91 (m, 5H,H-2, H-3, H-4, H-5, H-6), 2.12-1.90 (m, 1H, thp-ring), 1.85-1.32 (m; 5H,thp-ring)

[0536]³¹P-NMR (81 Hz, D₂O): δ 2.13 (1P), 1.85 (1P), 0.95 (2P)

[0537]¹³C-NMR (50.3 MHz, D₂O): δ 103.8 (1C, OCH₂O), 80.7 (1C), 80.0(1C), 77.7 (1 C), 76.5 (2 C), 30.3 (1C), 29.6 (1C), 15.5 (1C)

[0538] MS (neg. mode, FAB, glycerol): 573 [M−2H⁺+Na⁺] (13), 551 [M−H⁺](30)

[0539] Synthesis of (1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis[[bis(acetoxymethoxy)-phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.0⁶1-dodecane(8)

[0540] 7 (0.127 g, 0.23 mmol) was thoroughly dried and dissolved in 2.5mL dry acetonitrile. Diisopropylethyl amine (0.28 mL, 1.65 mmol) andacetoxymethyl bromide (0.164 mL, 1.64 mmol) were added and the solutionwas stirred for 48 hours at room temperature in the dark. All volatilecomponents were removed in high vacuum and the oily residue was purifiedby preparative HPLC (RP18, 40 mL/min, 60% MeOH, t_(Ret)=20.25 min). 8was isolated in 33% yield (0.045 g, 0.043 mmol) as a clear oil.

[0541]¹H-NMR (200 MHz, d₈-toluene): δ 5.91-5.55 (m, 16H, OCH₂OAc), 5.42(br s, 1H, OCH₂O), 5.17-5.00 (m, 2H) and 4.96-4.76 (m, 4H H-2, H-3, H-4,H-5), 4.25 (d, 1H, ³J=4.3 Hz, H-6), 2.31-2.13 (m, 2H, thp-ring), 1.88(s, 3H, CH₃), 1.88 (s, 3H, CH₃), 1.87 (s, 3H, CH₃), 1.86 (s, 9H, 3*CH₃),1.84 (s, 3H, CH₃), 1.76 (s, 3H, CH₃), 1.59-1.22 (m, 4H, thp-ring)

[0542]¹³C-NMR (50.3 MHz, d₈-toluene): δ 69.34-168.82 (C), 169.22 (C),169.13 (C), 168.85 (C), 104.24 (CH₂), 83.69-82.96 (CH₂), 81.66 (C),79.34 (CH), 78.50 (CH), 78.21 (CH), 77.20 (CH), 77.03 (CH), 30.28 (CH₂),30.03 (CH₂), 21.87-19.90 (CH₃—Ac), 16.00 (C—H₂)

[0543]³¹P-NMR (81 MHz, d₈-toluene): δ 2.72 (1P), −2.75 (1P), −3.36(1P),−3.42 (1P)

[0544] MS (neg. mode, FAB, NBA) in/z (%): 1055 [M−CH₂OAc⁺] (45), 983[M−2CH₂OAc⁺+H⁺] (7)

[0545] MS (pos. mode, FAB, NBA) m/z (%): 1151 [M+Na⁺] (9), 1129 [M+H⁺](18), 1057 [M+2H⁺−CH₂OAc⁺] (24), 985 [M+3H⁺−2CH₂OAc⁺] (26), 913[M+4H⁺−3CH₂OAc⁺] (24), 841 [M+5H⁺−4CH₂OAc⁺] (24), 769[M+6H⁺−5CH₂OAc^(+] ()22), 697 [M+7H⁺6CH₂OAc⁺] (18), 621 [M+8H⁺−7CH₂OAc⁺](5)

[0546] HR-MS calcd. for C₃₁H₄₇O₃₂P₄ [M−CH₂OAc⁺] 1055.1001, found1055.1029

EXAMPLE 8 Cystic Fibrosis Human Nasal Epithelial Cell Assay

[0547] CF Human Nasal Epithelial (CFHNE) Cell Isolation andProliferation

[0548] Nasal Polyps were surgically obtained from a CF patient atChildren's Hospital (Seattle, Wash.), transported on ice in a sterilecontainer containing a 1:1 mixture of Dulbecco's modification of minimumessential medium Eagle and Ham's F-12 nutrient medium (DMEM/F-12)(Irvine Scientific, Santa Ana, Calif.) supplemented with 100 U/mLpenicillin, 0.1 mg/mL streptomycin, 10 mM HEPES, and 2 mM L-glutamine.The tissue samples were aseptically removed from the transport mediumand washed (repeated 5 times) by suspending in 40 mL of Joklik'smodification of minimum essential medium Eagle (JMEM) at 4° C., andcentrifuging at 500 rpm. The supernatant was aspirated and discarded.The tissue was then transferred to JMEM containing 200 U/mL penicillin,0.2 mg/mL streptomycin, 0.1 mg/mL gentamycin sulfate (Clonetics, SanDiego, Calif.), and 0.1 μg/mL amphotericin-B (Clonetics), and 0.1%Protease, washed an additional two times, suspended in 15 nm in a 10 cmtissue culture dish, and incubated at 4° C. for 24 hours. The tissuesamples were then gently triturated, the connective tissue asepticallyremoved, and the remaining cell suspension centrifuged at 1000 rpm. Thesupernatant was aspirated and the pellet was resuspended in 10 mLproliferation media, consisting of Keratinocyte-Serum Free Medium (KSFM)(Gibco-BRL, Grand Island, N.Y.) containing 5 ng/ml EGF (Gibco), 50 μg/mLBPE (Gibco), 5 μg/mL insulin, 5 μg/mL transferrin, 5 μg/ml sodiumselenite, 10 nM 3,3′,5-triiodo-L-thyronine, 200 nM hydrocortisone, 100U/mL penicillin, 0.1 mg/mL streptomycin, and 2 mM L-glutamine. The cellsuspension was transferred to 2.10 cm tissue culture dishes coated with3 μg/cm² rat-tail collagen (Type I) (Becton-Dickinson, Bedford, Mass.),incubated at 37° C. in an humidified atmosphere of 5% CO₂ and 95% air.The cells were allowed to grow for 6 days (˜70% confluence) with themedia being replaced with fresh media every other day. The cells werethen trypsinized using 0.025% trypsin-EDTA for 5 min. The cellsuspension was collected, the trypsin deactivated with 10% Fetal BovineSerum, and centrifuged at 1000 rpm for 5 min. The cells were thencounted using an hemocytometer with a yield of 3×10⁶ cells per dish. Thesupernatant was aspirated and the cells were resuspended in KSFM andplated at a density of 10⁴ cells/cm².

[0549] CFHNE Cell Ussing Chamber Preparation

[0550] The CFHNE cells were prepared for Ussing Chamber studies usingSnapwell permeable supports (0.4 μm pore size) (Corning Costar,Cambridge, Mass.) coated with 3 μg/cm² rat-tail collagen (type 1). Cellswere plated at 9×10³ cells/cm² in KSFM. After 2 days, an air-liquidinterface (ALI) was initiated in DMEM/F-12 containing: 2% Fetal CloneIII (Hyclone, Logan, Utah), 2.5 μg/mL insulin, 2.5 μg/mL transferrin,2.5 μg/mL sodium selenite, 20 nM hydrocortisone, 1.5 μM epinephrine, 50μg/mL BPE, 1 nM 3,3′,5-triiodo-L-thyronine, 3.3 nM All-trans retinoicacid, 250 nM ethanolamine, 250 nM O-phosporylethanolamine, 100 U/mLpenicillin, 0.1 mg/ml streptomycin, and 2 mM L-glutamine. The CFHNEmonolayers reached maximal resistance (˜200 Ω/cm²) 5-7 days afterinitiation of ALI. All media/reagents, unless otherwise specified, weresupplied by Sigma Chemical Co., St. Louis, Mo.

[0551] Effect of rac-2-O-Butyryl-2-C-propyl-1-O-propyl-myo-inositol3,4,5,6-Tetrakis-phosphate Octakis(acetoxymethyl) Ester

[0552] The compound rac-2-O-butyryl-2-C-propyl-1-O-propyl-myo-inositol3,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester (“Compound A”) wastested as described above. The results are shown in FIGS. 12A and 12B.As can be seen in FIG. 10A, Compound A (200 μM), added basolaterally,elicited a prolonged increase in I_(sc) comparable in amplitude to thatstimulated with ATP. Although 95 μM had little direct effect, itappeared to prolong the response to subsequent stimulation with ATP(FIG. 10B.)

CONCLUSION

[0553] Ins(3,4,5,6)P₄ is an endogenous inhibitor of Ca²⁺ activated Cl⁻(CLCA) channels. Ins(3,4,5,6)P₄ antagonists described herein overcomeinhibition and promote Cl⁻ secretion in human colonic epithelia.Rationale modification of the molecular structure of the antagonists hasyielded a 10-fold improvement in potency. In addition, some compoundswere generated that triggered transepithelial currents in primary nasalepithelia from CF patients and colonic epithelia. In at least one case,a chloride current was identified in CFHNE using patch clamp techniques.

EXAMPLE 9 Colonic Epithelial Cell Assay

[0554] Transepithelial chloride secretion is key to the regulation ofwater flow across gut epithelia. Diseases of chloride and fluidtransport include cystic fibrosis and secretory diarrhea. As theendogenous factors which regulate chloride secretion are uncovered, thechances that we will be able to better modulate fluid secretion indiseased epithelial is improved.

[0555] The Regulation of Cl⁻ Secretion in Colonic Epithelia.

[0556] Transepithelial chloride secretion in colonic epithelia iscontrolled through at least three separate apically located chloride Cl⁻channels and a least two separately regulated basolateral potassiumchannels. Apical chloride efflux is counterbalanced by basolateral Kefflux, preserving electroneutrality. Two of the apical chloridechannels, the cystic fibrosis transmembrane regulator and the outwardlyrectifying chloride channel are functionally linked and triggered byelevations in cAMP levels. The calcium dependent chloride channel isregulated independently of the cAMP triggered pathways. There are twomain signaling pathways regulating this function T₈₄ monolayers retainmany receptor-mediated Cl⁻ secretory mechanisms including thoseinvolving changes in free cytosolic Ca²⁺ ([Ca²⁺]_(i)) and the CFTR.Agents which elevate [Ca²⁺]_(i) such as histamine, carbachol, Ca²⁺ionophores and thapsigargin, stimulate varying degrees of Cl⁻ secretionacross T₈₄ monolayers (Dharmsathaphorn et al., “Multiplecalcium-mediated effector mechanisms regulate chloride secretoryresponses in T84 cells,” Am. J. Physiol. 256:C1224-C₁₂₃₀ (1989);Kachintorn et al., “Activation by calcium alone of chloride secretion inT84 epithelial cells” British J: Pharmacology 109:510-517 (1993)). Arise in [Ca²⁺]_(i) can trigger Cl⁻ secretion through effects on both Cl⁻and K⁺ channels, since basolateral K efflux can be rate limiting becauseit is necessary for the restoration of electroneutrality. Althoughcyclic AMP also stimulates Cl⁻ secretion in T₈₄ cells, Ca²⁺ mediated Cl⁻secretion (CaMCS) is through a Cl⁻ channel which is distinct from thecyclic AMP mediated Cl⁻ secretion and is modulated by the Ca/calmodulindependent protein kinase II (CaMKII) (Worrel et al., “CaMKII mediatesstimulation of chloride conductance by calcium in T84 cells,” Am. J:Physiol. 260(Cell Physiol 29):C877-C882 1991; Preston, et al.,“Calmodulin defects cause the loss of Ca⁺⁺-dependent K⁺ currents in twopantophobiac mutants of Paramecium tetraurelia,” J. Memb. Biol.115:51-60, (1990); Okada, et al., “Evidence for the involvement ofcalmodulin in the operation of Ca⁺⁺-activated K⁺ channels in mousefibroblasts,” J. Memb. Biol. 96:121-128 (1987).

[0557] Role of Inositol Polyphosphates in the Regulation of ChlorideSecretion.

[0558] We have found that the endogenous regulator of Ca²⁺-mediated Cl⁻secretion, D-myo-inositol(3,4,5,6)tetrakisphosphate (Ins(3,4,5,6)P₄),becomes elevated in T₈₄ cells following stimulation with cholinergicagonists but not histaminergic agonists or cyclic AMP-elevating agents.(Kachintorn, et al. 1992). Elevation of D-myo-Ins(3,4,5,6)P₄ levels,either through cholinergic stimulation or addition of a cell permeantform of Ins(3,4,5,6)P₄ results in the uncoupling of subsequent Cl⁻secretion from the elevation of intracellular Ca²⁺ levels (Vajanaphnich,et al., “Long-term uncoupling of chloride secretion from intracellularcalcium levels by Ins(3,4,5,6)P₄ ,” Nature 371:711-714 (1994)). Studiesusing whole cell patch clamp with intracellular perfusion of inositoltetrakisphosphates indicate that Ins(3,4,5,6)P₄ directly modulates anapically located Ca²⁺/calmodulin kinase regulated Cl⁻ channel in T₈₄(Xie, et al., J. Biol. Chem. 271:14092-14097 (1996)). This effect may bemediated by an okadaic acid sensitive protein phosphatase. On the otherhand, Ca²⁺ dependent Cl⁻ channels reconstituted in planar lipid bilayerscould also be regulated by low levels of Ins(3,4,5,6)P₄ in the absenceof phosphatase activities (Ismailov, et al., “A biologic function for an“orphan” messenger: D-myo-inositol 3,4,5,6-tetrakisphosphate selectivelyblocks epithelial calcium-activated chloride channels,” Proc Natl AcadSci USA 93:10505-9 (1996).

[0559] Method Detail:

[0560] Cl⁻ secretion in colonic epithelial cells was measured as shortcircuit current (I_(sc)) across T₈₄ monolayers grown to confluence onSnap-wells (Costar) and mounted in Ussing chambers equipped with voltageclamps (Physiologic Instruments, San Diego). Data was acquired andanalyzed using AAcquire and Analyze@ software (Physiologic Instruments).T₈₄ cells (passages 18-48) were maintained as described previously inMcRoberts, J. A. et al., “Cyclic AMP and Ca²⁺-activated K⁺ transportpathway in the apical membrane of a human colonic epithelial cell line,”J. Biol. Chem. 260:14163-14172 (1985). After 0.7-10 days of incubationon Snap-wells, monolayers had formed tight junctions and werepreincubated with 100, 200 or 400 mM Bt₂D-myo-Ins(3,4,5,6)P₄/AM for30-min. prior to mounting. An extracellular concentration of 200 uMcorresponds to an intracellular concentration of 4 uM (Vajanaphnich, etal., “Long-term uncoupling of chloride secretion from intracellularcalcium levels by Ins(3,4,5,6)P₄ ,” Nature 371:711-714 (1994)). After anadditional 30 min, 0.1 mM histamine was added and the I_(sc) monitored.Ca²⁺-dependent Cl⁻ secretion was stimulated with histamine, aphysiologically relevant agonist which does not have a measurable longterm effect on inositol polyphosphate metabolism as does carbachol inthis cell line (Kachintorn, et al., “Elevation of inositoltetrakisphosphate parallels inhibition of calcium-dependent chloridesecretion in T84 colonic epithelia cells,” Am. J. Physiol., Cell264:C671-C676 (1993); Kachintorn et al., “Activation by calcium alone ofchloride secretion in T84 epithelial cells” British J. Pharmacology109:510-517 (1993)).

[0561] Effects of 2,3,5-Tri-O-butyryl-myo-inositol 1,4,6-TrisphosphateHexakis-(proprionoxymethyl) Ester

[0562] The increase in short circuit current (I_(sc)) in monolayers ofT₈₄ cells in Ussing chambers exposed to differing concentrations of2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(proprionoxymethyl)ester (“Bt₃Ins(1,4,6)P₃/PM” or “TMX”) is shownin FIG. 9A (where ent-TMX is the enantiomer of2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(proprionoxymethyl)ester (TMX). FIG. 9A depicts the effects ofvarious doses of TMX on short circuit current (I_(sc)) in colonic andairway epithelial cells. This effect is specific because the enantiomerof TMX (ent-TMX-1,2,5-tri-O-butyryl-myo-inositol 3,4,6-trisphosphatehexakis-(proprionoxymethyl) ester is inactive (FIGS. 11A and 11C). Thepro-secretory activity of TMX may be attributed, at least in part, to astimulatory effect on intracellular calcium levels due to its structuralsimilarity to inositol 1,4,5-trisphosphate. In addition, its inhibitoryeffect on the accumulation of Ins(3,4,5,6)P₄ would enableCa²⁺-stimulated Cl⁻ secretion. In addition, it may directly trigger Cl⁻secretion through binding to Cl⁻ channels directly.

[0563] Effect of rac-1,2-O-Isopropylidene-2-C-propyl-myo-inositol3,4,5,6-Tetrakisphosphate octakis(acetoxymethyl) Ester (“Compound B”)

[0564] Using the foregoing procedures, the compoundrac-1,2-O-isopropylidene-2-C-propyl-myo-inositol3,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester (“Compound B”) wastested and compared with TMX. FIG. 11 shows the effect of TMX andrac-1,2-O-isopropylidene-2-C-propyl-myo-inositol3,4,5,6-tetrakis-phosphate octakis(acetoxymethyl)ester (“Compound B”) onthe negative component of the ATP effect on I_(sc) in CFHNE. As shown inFIG. 12, Compound B triggers an increase in I_(sc) in T₈₄ cells.Compound B was applied acutely to the apical surface of T₈₄ monolayersmounted in Ussing chambers at the indicated time. After 25 min. 10⁻⁴ Mcarbachol was added. Data plotted as ΔI_(sc) (μA/cm²) in FIG. 12represents one monolayer. As shown in FIG. 12, ATP (10⁻⁴ M) stimulated atransient increase in I_(sc) followed by a prolonged decrease when addedto monolayers pretreated with vehicle. This decrease was attenuated inmonolayers exposed to TMX for 10 min. In contrast, althoughpreincubation with, 200 μM Compound B(rac-1,2-O-isopropylidene-2-C-propyl-myo-inositol3,4,5,6-tetrakisphosphate octakis(acetoxy-methyl) ester) inhibited thetransient increase following ATP, it totally reversed the negativephase, instead causing a long term increase in I_(sc) in CFHNEmonolayers. Compound B also had an acute effect in T₈₄ monolayers,triggering a large and prolonged increase in I_(sc).

[0565] Effect of rac-1,2-O-Isopropylidene-2-C-propyl-myo-inositol3,4,5,6-Tetrakis-phosphate octakis(acetoxymethyl) Ester (“Compound B”)At 400 μM with 30 Minute Preincubation

[0566] We have previously demonstrated that carbachol appliedbasolaterally to the monolayers of T₈₄ colonic epithelia for a period of5 minutes or more results in elevated intracellular levels ofIns(3,4,5,6)P₄ that mediates inhibition of subsequent increase in I_(sc)stimulated by the calcium-elevating agent, thapsigargin (Vajanaphanich,et al. 1994 Nature). Ins(3,4,5,6)P₄ elevation may also mediate the “turnoff” of the increased I_(sc) following carbachol. Inositol polyphosphateanalogs of Ins(3,4,5,6)P₄ with substitutions at the 1 and 2 positionthat restrict the flexibility of the molecule could compete withendogenous Ins(3,4,5,6)P₄-mediated inhibition and prolong and augmentchloride secretion stimulated through elevation of intracellularcalcium. For instance, preincubation with Compound B for 30 minutes bothprolongs the carbachol induced increase in I_(sc) in colonic epithelia,T₈₄, and overcomes the inhibitory effect of carbachol pretreatment onthe thapsigargin induced increase in I_(sc) as shown in FIG. 13.

[0567] Effect of(5aa,6a,7b,8a,9b,9aa)-6,7,8,9-Tetrakis[[bis(acetoxymethoxy)phosphoryl]-oxy]-5a-[(2′-methoxy)ethoxy]methoxy-decahydro-benzo[b]oxepin(“Compound C”) At 200 μM with 30 Minute Preincubation

[0568] Monolayers of T₈₄ colonic epithelia were preincubated with 200 μMor 400 μM Compound C for 30 minutes prior to mounting in Ussingchambers. The I_(sc) response to stimulation with the calcium elevatingagents carbachol and thapsigargin were subsequently compared anddepicted in FIGS. 14 and 15. Both concentrations of Compound C inhibitedthe response to carbachol by approximately 50% as seen in FIGS. 14 and15.

[0569] Effect of rac-2-O-Butyryl-2-C-propyl-1-O-propyl-myo-inositol3,4,5,6-Tetrakisphosphate-octakis(acetoxymethyl)ester (“Compound A”) At200 μM with 30 Minute Preincubation (Carbachol and Thapsigargin)

[0570] Monolayers of T₈₄ colonic epithelia were preincubated with 400 μMCompound A for 30 minutes prior to mounting in Ussing chambers. TheI_(sc) response to stimulation with the calcium elevating agentscarbachol and thapsigargin were subsequently compared and depicted inFIG. 16. Pretreatment with Compound A did not affect the response tocarbachol but partially reversed the inhibition of thapsigargin inducedchloride secretion as seen in FIG. 16.

[0571] Effect of rac-1-O-butyryl-2-C-propyl-myo-inositol 1,2,4,5,6pentakisphosphate-dekakis(acetoxymethyl) ester (Compound D) on I_(sc) ina human colonic epithelial cell line, T₈₄.

[0572] Monolayers of the human colonic epithelial cell line, T₈₄ weremounted in Ussing chambers and treated acutely with Compound D. As canbe seen in FIG. 17 200 μM Compound D stimulated a small increase inbasal I_(sc) but had little effect on subsequent carbachol-stimulatedI_(sc).

[0573] Effect of (4aa, 5a,6b,7a,8b,8aa)-5,6,7,8-tetrakis[bis9acetoxymethoxy)phosphoryl]-oxy]-4a-butyryloxy-octahydro-chromen(Compound G) At 200 μM with 30 Minute Preincubation (Carbachol andThapsigargin)

[0574] Monolayers of T₈₄ colonic epithelia were preincubated with 200 μMCompound G for 30 minutes prior to mounting in Ussing chambers. TheI_(sc) response to stimulation with the calcium elevating agentscarbachol and thapsigargin were subsequently compared and depicted inFIG. 18. Pretreatment with Compound G did not affect the response tocarbachol but reversed the inhibition of thapsigargin induced chloridesecretion as seen in FIG. 18.

EXAMPLE 10 Inflammation Studies

[0575] Materials:

[0576] Arachidonic Acid (Nu-Chek, Elysian, Minn. 56028); Indomethacin(stock=12.5 mg/mL acetone). Prednisolone (Sigma Chem, P6004) 20 mMdissolved in acetone; Animals: Female Balb/c mice (B&K laboratories) andSwiss-Webster hybrid (B& K Laboratory, Kent, Wash.)

[0577] Method

[0578] The right ear of each mouse was used for the test compound.Unless otherwise indicated each compound was adjusted to 20 mM bydilution into DMSO or DMSO/5% Pluronic F127 as indicated and 1 or 2microliters were applied to the test ear. Thirty minutes after eachcompound was applied, 4 mg arachidonic acid was applied (4 μL/ear; 2μL/side) to both ears of each mouse. Measurements of edema were taken 60min after the application of arachidonic acid with microcalipers(Oditest, Scientific Supply and Equipment) unless otherwise indicated.The results are shown in FIG. 15 as millimeters×10⁻² measured at thethickest point of each ear. Control measurements were performed on earsthat were only treated with arachidonic acid.

[0579] Response to prednisolone (20 μL of 20 mM/ear; 10 μl/side) added 3hrs prior 20′ to arachidonic acid or indomethacin (5 μg/ear) added 30min prior to test compounds was determined. The results are shown inTable A, where the anti-inflammatory/inflammatory effects of 20nanomoles/ear of the indicated compounds were tested as described inMethods. Each plus represents a change from control (arachidonic acidalone) of 2.5 mm×10⁻². Effects were compared with those of Indomethacin(400 nmols/ear). TABLE A Effects of Inositol Polyphosphate Analogs onarachidonic acid-induced inflammation in mouse ears Ear ThicknessCompound¹ (mm × 10⁻²) n Control (arachidonic 56.5 ± 1.57 10  acid +vehicle) A 52.5 ± 5.5 3 B 56.1 ± 2.1 3 C 50.0 ± 5.5 3 D 44.0 ± 1.4 4 D44.2 ± 6.5 3 Tri-cyclo 49.8 ± 2.5 4 TMX/AM 44.1 ± 2.3 3

[0580] Acute inflammation: Membrane permeant inositol polyphosphateanalogs were administered topically and tested for inhibition ofinflammatory symptoms induced by arachidonic acid on the ear of SwissWebster mice. D-2,3-Di-O-butyryl-myo-inositol 1,4,5,6 tetrakisphosphateoctakis(acetoxymethyl)ester had an inhibitory effect comparable inintensity to that of indomethacin (250 μg/animal) at a dose of 20-30μg/animal. More potent was D-2,3-Di-O-butyryl-myo-inositol 1,4,5,6tetrakisphosphate octakis(propionoxymethyl)ester which had topicalanti-inflammatory effects with values ranging from 0.25-1.5 μg/animal.D-1-O-octyl-2-O-butyryl-myo-inositol1,4,5,6-tetrakisphosphate-octakis-(propionoxymethyl)ester (INO-E3) wasyet more potent with an ED₅₀ of 7-14 :g/kg, and has been shown to beeffective when applied topically, intraperitoneally and orally. Nomacroscopic negative effects were observed with ip injections up to 50μg/animal and iv injections of 20 μg/animal several weeks aftertreatment.

[0581] Several modes of delivery were assayed, including a simple cream.Systemic effects were observed with more lipophilic formulations.Compounds injected into the tail vein of the mouse or delivered ipresulted in an anti-inflammatory effect comparable to the topicalapplications, with no apparent side effects. Therefore, inositolpolyphosphate derivatives show promise as both topical and systemicanti-inflammatory agents for the treatment of diseases such asinflammatory bowel disease, arthritis, asthma, and adult respiratorydistress syndrome in addition to contact dermatitis.

REFERENCES

[0582] 1. Carlson, R P. O'Neill-Davis, L. Chang, J. Lewis, A J. 1985.Modulation of mouse ear edema by cyclooxygenase and lipoxygenaseinhibitors and other pharmacologic agents. Agents and Actions, 17:2,197-204.

[0583] 2. Chang, J. Blazek, E. Skowronek, M. Marinari, L. Carlson, R P.1987. The antiinflammatory action of guanabenz is mediated through5-lipoxygenase and cyclooxygenase inhibition. Europ. J. Pharmac. 142,197-205

[0584] 3. Simon, L S, MD 1999. Role and regulation of Cyclooxygenase-2during inflammation. Am. J. Med, 106(5B): 37S-42S

EXAMPLE 11 Composition for Systemic Administration for Lung Pancreas,Gastrointestinal and Vaginal and Ductile Manifestations of CysticFibrosis

[0585] A composition comprising physiological saline and atherapeutically effective amount of an inositol derivative of Examples1-7 is administered intravenously to a patient on a daily prophylacticbasis. As a result of the administration, the patient has reducedviscosity of mucosal secretions, improved airway clearance andoxygenation of the blood.

EXAMPLE 12 Composition for Patient with Intestinal Symptoms

[0586] A composition comprising a physiologically acceptable carrier orvehicle and a therapeutically effective amount of an inositol derivativeof Examples 1-7 the present invention is administered once daily for oneweek, in the form of an enema, to a patient having ulcerative colitis.As a result of the administration, the patient has improved digestion,reduced risk of intestinal obstruction and improved regularity of bowelfunction.

[0587] While the preferred embodiment of the invention has beenillustrated and described, it will be appreciated that various changescan be made therein without departing from the spirit and scope of theinvention.

The embodiments of the invention in which an exclusive property orprivilege is claimed ate defined as follows:
 1. A compound of theformula:

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain, alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃ isindependently selected from a group consisting of hydrogen and —C₁-C₁₂alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, and benzyl,said R₃, except hydrogen, being unsubstituted or substituted with one ormore halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, and OC(O)C₁-C₆ alkylgroups; and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.
 2. A compound of claim 1 selected from the groupconsisting of D-2,3,5-tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(acetoxymethyl)ester, D-2,3,5-Tri-O-butyryl-myo-inositol1,4,6-trisphosphate hexakis(propionoxymethyl) ester, and the racemates,diastereomers and pharmaceutically acceptable salts thereof.
 3. Acompound of the formula:

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃ isindependently selected from a group consisting of hydrogen and —C₁-C₁₂alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, and benzyl,said R₃, except hydrogen, being unsubstituted or substituted with one ormore halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, and OC(O)C₁-C₆ alkylgroups, including racemates and diastereomers thereof, with the provisothat the compound of formula (II) is not D-3,6-Di-O-butyryl-myo-inositol1,2,4,5-tetrakisphosphate octakis(acetoxymethyl) ester; orD-1,4-Di-O-butyryl-myo-inositol 2,3,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester; and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 4. A compound of the formula:

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;or each R₁ may be taken together to form a cyclic structure; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain alkyl and each R₃ isindependently selected from a group consisting of —C₁-C₂₀ straight orbranched chain alkyl, —C₂-C₂₀ straight or branched chain alkenyl oralkynyl, —C₁-C₁₁—X—Y, where each X is independently selected from agroup consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—, —CO(S)—, —OC(O)—,—SC(O)—, OC(S)—, or —NH—, and where Y is independently selected from agroup consisting of —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂straight or branched chain alkenyl or alkynyl, or unsubstituted andsubstituted benzyl, and phenyl; and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 5. A compound of claim 4selected from the group consisting of rac-(3aS, 4R, 5S, 6R, 7S,7aS)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol;rac-(1S, 2R, 3S, 4R, 5S,6S)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane;rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxy-methoxy)phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane;and the racemates, diastereomers and pharmaceutically acceptable saltsthereof.
 6. A compound of the formula:

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain allyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;each R₂ is independently selected from a group consisting of hydrogenand —C(R³)(R³)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃is independently selected from a group consisting of hydrogen and—C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring, phenyl, andbenzyl, said R₄, except hydrogen, being unsubstituted or substitutedwith one or more halogen OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, andOC(O)C₁-C₆ alkyl groups, including racemates and diastereomers thereof;and each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, and —C(X)(Y)—C(X)(Y)—C(X)(Y)—; where X andY are independently selected from a group consisting of hydrogen,—C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight or branchedchain alkenyl or alkynyl, or unsubstituted and substituted benzyl, andphenyl, including racemates and diastereomers thereof and hydrogen, —OH,—Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl, —CO(S)alkyl,—OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or —CONHalkyl, or—CON(alkyl)(alkyl), —NH₂; and n is an integer from 2 to 13, preferably 2to 5; and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.
 7. A compound of the formula:

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;each R₂ is independently selected from a group consisting of hydrogenand —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃is independently selected from a group consisting of hydrogen and—C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, andbenzyl, said R₃, except hydrogen, being unsubstituted or substitutedwith one or more halogen, —OH, C₁-C₆ alkyl, —NO₂, —OC₁-C₆ alkyl, and—OC(O)C₁-C₆ alkyl groups, including racemates and diastereomers thereof;and each V is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)—; and m is aninteger from 2 to 15, preferably from 2 to 5; and each W isindependently selected from a group consisting of —C(X)—,—C(X)—C(X)(Y)—, or —C(X)—C(X)(Y)—C(X)(Y)—; and n is an integer from 1 to15, preferably from 1 to 3; and X and Y are independently selected froma group consisting of hydrogen, —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof and hydrogen, —OH, —Oalkyl, —SH, —Salkyl,—CO(O)alkyl, —CS(O)alkyl, —CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl,—OC(S)alkyl, or —CONH₂, or —CONHalkyl, or —CON(alkyl)(alkyl), —NH₂; andthe racemates, diastereomers and pharmaceutically acceptable saltsthereof.
 8. A method for regulating chloride secretion and/or inhibitinginflammation in a cell or tissue, comprising contacting the cell ortissue with an effective amount of a compound of formula (I):

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight- orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃ isindependently selected from a group consisting of hydrogen and —C₁-C₁₂alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, and benzyl,said R₃, except hydrogen, being unsubstituted or substituted with one ormore halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, and OC(O)C₁-C₆ alkylgroups; and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.
 9. A method of claim 8, wherein the compound is selectedfrom the group consisting of D-2,3,5-tri-O-butyryl-myo-inositol1,4,6-trisphosphate hexakis-(acetoxymethyl) ester;D-2,3,5-Tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(propionoxymethyl)ester, and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 10. A method of regulatingchloride secretion and/or inhibiting inflammation in a human or animalpatient in need thereof comprising administering to the patient atherapeutically effective amount of a compound of the formula:

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃ isindependently selected from a group consisting of hydrogen and —C₁-C₁₂alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, and benzyl,said R₃, except hydrogen, being unsubstituted or substituted with one ormore halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, and OC(O)C₁-C₆ alkylgroups; and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.
 11. A method of claim 10, wherein the compound isselected from the group consisting of D-2,3,5-tri-O-butyryl-myo-inositol1,4,6-trisphosphate hexakis-(acetoxymethyl) ester;D-2,3,5-Tri-O-butyryl-myo-inositol 1,4,6-trisphosphatehexakis(propionoxymethyl)ester, and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 12. A method for regulatingchloride secretion and/or inhibiting inflammation in a cell or tissue,comprising contacting the cell or tissue with an effective amount of acompound of formula (II):

each R₁ is independently selected from the group consisting of hydrogen,—OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀ straight orbranched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight or branchedchain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and —OC₂-C₂₀straight or branched chain alkenyl or alkynyl; each R₂ is independentlyselected from a group consisting of hydrogen and —C(R₃)(R₃)OC(O)C₁-C₄straight or branched chain alkyl; and each R₃ is independently selectedfrom a group consisting of hydrogen and —C₁-C₁₂ alkyl, both R₃ taken asa 5- or 6-membered ring, phenyl, and benzyl, said R₃, except hydrogen,being unsubstituted or substituted with one or more halogen, OH, C₁-C₆alkyl, —NO₂, —OC₁-C₆ alkyl, and OC(O)C₁-C₆ alkyl groups; and theracemates, diastereomers and pharmaceutically acceptable salts thereof.13. A method of claim 12, wherein the compound is selected from thegroup consisting of D-3,6-di-O-butyryl-myo-inositol1,2,4,5-tetrakisphosphate octakis(propionoxymethyl)ester,D-1,4-di-O-butyryl-myo-inositol 2,3,5,6-tetrakis-phosphateoctakis(propionoxymethyl)ester, and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 14. A method of regulatingchloride secretion and/or inhibiting inflammation in a human or animalpatient in need thereof comprising administering to the patient atherapeutically effective amount of a compound of the formula:

each R₁ is independently selected from the group consisting of hydrogen,—OH, —C₁-C₂₀ straight or branched chain allyl, —C₂-C₂₀ straight orbranched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight or branchedchain alkyl and —OC₁-C₂₀ straight or branched chain alkyl, and —OC₂-C₂₀straight or branched chain alkenyl or alkynyl; each R₂ is independentlyselected from a group consisting of hydrogen and —C(R₃)(R₃)OC(O)C₁-C₄straight or branched chain alkyl; and each R₃ is independently selectedfrom a group consisting of hydrogen and —C₁-C₁₂ alkyl, both R₃ taken asa 5- or 6-membered ring, phenyl, and benzyl, said R₃, except hydrogen,being unsubstituted or substituted with one or more halogen, OH, C₁-C₆alkyl, —NO₂, —OC₁-C₆ alkyl, and OC(O)C₁-C₆ alkyl groups; and theracemates, diastereomers and pharmaceutically acceptable salts thereof.15. A method of claim 14, wherein the compound is selected from thegroup consisting of D-3,6-di-O-butyryl-myo-inositol1,2,4,5-tetrakisphosphate octakis(propionoxymethyl)ester;D-1,4-di-O-butyryl-myo-inositol 2,3,5,6-tetrakis-phosphateoctakis(propionoxymethyl)ester, and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 16. A method for regulatingchloride secretion and/or inhibiting inflammation in a cell or tissue,comprising contacting the cell or tissue with an effective amount of acompound of formula (III):

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain al. V, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;or each R₁ may be taken together to form a cyclic structure; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃ isindependently selected from a group consisting of —C₁-C₂₀ straight orbranched chain alkyl, —C₂-C₂₀ straight or branched chain alkenyl oralkynyl, —C₁-C₁₁—X—Y, where each X is independently selected from agroup consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—, —CO(S)—, —OC(O)—,—SC(O)—, OC(S)—, or —NH—, and where Y is independently selected from agroup consisting of —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂straight or branched chain alkenyl or alkynyl, or unsubstituted andsubstituted benzyl, and phenyl, including racemates and diastereomersthereof. each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ al,and OC(O)C₁-C₆ alkyl groups; and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 17. A method of claim 16wherein the compound is selected from the group consisting of rac-(3aS,4R, 5S, 6R, 7S,7aS)4,5,6,7-tetrakis[[bis(acetoxy-methoxy)phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol;rac-(1S, 2R, 3S, 4R, 5S,6S)-4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane;rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxymethoxy)phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane;and the racemates, diastereomers and pharmaceutically acceptable saltsthereof.
 18. A method of regulating chloride secretion and/or inhibitinginflammation in a human or animal patient in need thereof comprisingadministering to the patient a therapeutically effective amount of acompound of the formula (III):

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;or each R₁ may be taken together to form a cyclic structure; each R₂ isindependently selected from a group consisting of hydrogen and—C(R₄)(R₄)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃ isindependently selected from a group consisting of —C₁-C₂₀ straight orbranched chain alkyl, —C₂-C₂₀ straight or branched chain alkenyl oralkynyl, —C₁-C₁₁—X—Y, where each X is independently selected from agroup consisting of —O—, —S—, —Se—, —CO(O)—, —CS(O)—, —CO(S)—, —OC(O)—,—SC(O)—, OC(S)—, or —NH—, and where Y is independently selected from agroup consisting of —C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂straight or branched chain alkenyl or alkynyl, or unsubstituted andsubstituted benzyl, and phenyl, including racemates and diastereomersthereof each R₄ is independently selected from a group consisting ofhydrogen and —C₁-C₁₂ alkyl, both R₄ taken as a 5- or 6-membered ring,phenyl, and benzyl, said R₄, except hydrogen, being unsubstituted orsubstituted with one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆alkyl, and OC(O)C₁-C₆ alkyl groups; and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 19. A method of claim 18wherein the compound is selected from the group consisting of rac-(3aS,4R, 5S, 6R, 7S,7aS)4,5,6,7-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-2,2-dimethyl-3a-propyl-hexahydrobenzo-(1,3)-dioxol;rac-(1S, 2R, 3S, 4R, 5S,6S)-4,5,6,7-tetrakis[[is(acetoxymethoxy)phosphoryl]oxy]-1-butyroxy-6-propoxy-1-propyl-cyclohexane;rac-(1S, 2R, 3S, 4R, 5S,6S)-2,3,4,5,6-pentakis-[[bis(acetoxy-methoxy)phosphoryl]oxy]-6-butyroxy-2-propyl-cyclohexane;and the racemates, diastereomers and pharmaceutically acceptable saltsthereof.
 20. A method for regulating chloride secretion and/orinhibiting inflammation in a cell or tissue, comprising contacting thecell or tissue with an effective amount of a compound of formula (IV):

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀′ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;each R₂ is independently selected from a group consisting of hydrogenand —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃is independently selected from a group consisting of hydrogen and—C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, andbenzyl, said R₃, except hydrogen, being unsubstituted or substitutedwith one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, andOC(O)C₁-C₆ alkyl groups, including racemates and diastereomers thereof;and each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, and —C(X)(Y)—C(X)(Y)—C(X)(Y)—; where X andY are independently selected from a group consisting of hydrogen,—C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight or branchedchain alkenyl or alkynyl, or unsubstituted and substituted benzyl, andphenyl, including racemates and diastereomers thereof and hydrogen, —OH,—Oalkyl, —SH, —Salkyl, —CO(O)alkyl, —CS(O)alkyl, —CO(S)alkyl,—OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or —CONHalkyl, or—CON(alkyl)(alkyl), —NH₂; and n is an integer from 2 to 13, preferably 2to 5; and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.
 21. A method of claim 20 wherein the compound is selectedfrom the group consisting ofD-2-O-butyryl-1,2,4,5,6-pentahydrobenzoxirane 1,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester;D-2-O-butyryl-2,3,4,5,6-pentahydrobenzoxirane 3,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester; D-2-O-butyryl1,2,4,5,6-pentahydrochroman1,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester;D-2-O-butyryl-2,3,4,5,6-pentahydrochroman 3,4,5,6-tetraakisphosphateoctakis(acetoxymethyl)ester; and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 22. A method of regulatingchloride secretion and/or inhibiting inflammation in a human or animalpatient in need thereof comprising administering to the patient atherapeutically effective amount of a compound of the formula (T):

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;each R₂ is independently selected from a group consisting of hydrogenand —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃is independently selected from a group consisting of hydrogen and—C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, andbenzyl, said R₃, except hydrogen, being unsubstituted or substitutedwith one or more halogen, OH, C₁-C₆ alkyl, NO₂, —OC₁-C₆ alkyl, andOC(O)C₁-C₆ alkyl groups, including racemates and diastereomers thereof;and each W is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, and —C(X)(Y)—C(X)(Y)—C(X)(Y)—; where X andY are independently selected from a group consisting of hydrogen,—C₁-C₁₁ straight or branched chain alkyl, —C₂-C₁₂ straight or branchedchain alkenyl or alkynyl, or unsubstituted and substituted benzyl, andphenyl, including racemates and diastereomers thereof and hydrogen, —OH,—Oalkyl, —SH, —S allyl, —CO(O)alkyl, —CS(O)alkyl, —CO(S)alkyl,—OC(O)alkyl, —SC(O)alkyl, —OC(S)alkyl, or —CONH₂, or —CONHalkyl, or—CON(alkyl)(alkyl), —NH₂; and n is an integer from 2 to 13, preferably 2to 5; and the racemates, diastereomers and pharmaceutically acceptablesalts thereof.
 23. A method of claim 22 wherein the compound is selectedfrom the group consisting ofD-2-O-butyryl-1,2,4,5,6-pentahydrobenzoxirane 1,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester;D-2-O-butyryl-2,3,4,5,6-pentahydrobenzoxirane 3,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester, D-2-O-butyryl1,2,4,5,6-pentahydrochroman1,4,5,6-tetrakisphosphate octakis(acetoxymethyl)ester;D-2-O-butyryl-2,3,4,5,6-pentahydrochroman 3,4,5,6-tetrakisphosphateoctakis(acetoxymethyl)ester; and the racemates, diastereomers andpharmaceutically acceptable salts thereof.
 24. A method for regulatingchloride secretion and/or inhibiting inflammation in a cell or tissue,comprising contacting the cell or tissue with an effective amount of acompound of formula (V):

wherein each R₁ is independently selected from the group consisting ofhydrogen, —OH, —C₁-C₂₀ straight or branched chain alkyl, —C₂-C₂₀straight or branched chain alkenyl or alkynyl, —OC(O)C₁-C₂₀ straight orbranched chain alkyl and —OC₁-C₂₀ straight or branched chain alkyl,—OC₂-C₂₀ straight or branched chain alkenyl or alkynyl, and —OPO(OR₂)₂;each R₂ is independently selected from a group consisting of hydrogenand —C(R₃)(R₃)OC(O)C₁-C₄ straight or branched chain alkyl; and each R₃is independently selected from a group consisting of hydrogen and—C₁-C₁₂ alkyl, both R₃ taken as a 5- or 6-membered ring, phenyl, andbenzyl, said R₃, except hydrogen, being unsubstituted or substitutedwith one or more halogen, —OH, C₁-C₆ alkyl, —NO₂, —OC₁-C₆ allyl, and—OC(O)C₁-C₆ alkyl groups, including racemates and diastereomers thereof;and each V is independently selected from a group consisting of—C(X)(Y)—, —C(X)(Y)—C(X)(Y)—, or —C(X)(Y)—C(X)(Y)—C(X)(Y)—; and m is aninteger from 2 to 15, preferably from 2 to 5; and each W isindependently selected from a group consisting of —C(X)—,—C(X)—C(X)(Y)—, or —C(X)—C(X)(Y)—C(X)(Y)—; and n is an integer from 1 to15, preferably from 1 to 3; and X and Y are independently selected froma group consisting of hydrogen, —C₁-C₁₁ straight or branched chainalkyl, —C₂-C₁₂ straight or branched chain alkenyl or alkynyl, orunsubstituted and substituted benzyl, and phenyl, including racematesand diastereomers thereof and hydrogen, —OH, —Oalkyl, —SH, —Salkyl,—CO(O)alkyl, —CS(O)alkyl, —CO(S)alkyl, —OC(O)alkyl, —SC(O)alkyl,—OC(S)alkyl, or —CONH₂, or —CONHalkyl; or —CON(alkyl)(alkyl), —NH₂; andthe racemates, diastereomers and pharmaceutically acceptable saltsthereof.
 25. A method of claim 24 wherein the compound is selected fromthe group consisting of (1S, 2R, 3S, 4R, 5S, 6S,8R)-2,3,4,5-tetrakis[[bis(acetoxymethoxy)phosphoryl]oxy]-7,12-dioxa-tricyclo-[6.3.1.0^(1,6)]-dodecane;and the racemates, diastereomers and pharmaceutically acceptable saltsthereof.